S1p3 antagonists

ABSTRACT

The present invention relates to antagonists of the S1P3 receptor formula (A) as herein described and pharmaceutical compositions thereof. The compounds of formula (A) are useful in the preparation of a medicament, in particular for the treatment of Alzheimer&#39;s disease.

The present invention relates to novel antagonists of the S1P3 receptor of formula (A) as herein described and to their pharmaceutical uses.

Such antagonists can be used for the treatment of inflammation-related diseases such as arthritis, fibrosis, inflammatory syndromes, atherosclerosis, vascular diseases, asthma, bradycardia, acute lung injury, lung inflammation, cancer, ocular hypertension, glaucoma, neuroinflammatory diseases, Sandhoff's disease, kidney ischemia-reperfusion injury, pain, diabetic heart disease and neurodegenerative diseases such as Alzheimer's disease, Parkinson's disease, Amyotrophic lateral sclerosis, Huntington's disease or Multiple Sclerosis.

BACKGROUND OF THE INVENTION

The S1P3 receptor gene encodes for a member of the endothelial differentiation gene (EDG) family of receptors widely present in central and peripheral human tissues (Rosen et al., 2009; Ishii et al., 2001). S1P₃ receptor (also called: EDG3; LPB3; S1PR3; EDG-3) belong to a class of five (S1P₁₋₅) seven-spanning membrane proteins belonging to the class of G-Protein-Coupled Receptors (GPCRs), whose natural ligand is the bioactive lipid sphingosine-1-phosphate (SIP) (Chun et al., 2002). S1P is involved in a large array of cellular responses modulating several physiological processes such as innate immunity, wound healing, vascular endothelial cell functions, inflammatory response and others (Ishii et al., 2004; Brinkmann, 2007; Rosen et al., 2009; Maceyka et al., 2012). S1P is intracellularly produced, with the direct role of secondary messenger (Spiegel and Milstien, 2003), and extracellularly exported acting to S1P cell membrane receptors as endogenous ligand.

The SIP receptors expressed in many apparatuses are able to trigger signalling through a variety of heterotrimeric G proteins, including Gi/o, G12/13, and Gq. In the S1P₁₋₅ receptors family, S1P₃ has been shown to be functionally relevant in several physiological processes such as regulations of heart rate, angiogenesis and vascular contraction (Forrest et al., 2004; Sanna et al., 2004; Marsolais and Rosen, 2009; Means and Brown, 2009; Murakami et al., 2010), in embryonic angiogenesis development (Kono et al., 2004) or as autophagy modulator (Taniguchi et al., 2012). The S1P₃ receptor is also deeply involved in immunological processes (Brinkmann V. (2009). Importantly, mice lacking S1P₃ receptor did not show evident abnormalities indicating a non-essential role of the receptor for a normal animal development (Ishii et al., 2001). As mentioned, SIP plays an important role as essential modulator of innate immunity and inflammation inducer. S1P once produced and released as signalling molecule by a wide range of cell types or even non-nucleated cells (e.g. platelets) (Pyne and Pyne, 2000) can exert an important role in inflammation. As introduced, together with the whole SIP receptor family, S1P₃ receptor system has been largely studied focusing on its role in disease and has been shown to be involved in a large number of pathologies. From the literature S1P₃ emerges as an important target implicated in pathologies with inflammatory components, in this case a pharmacological inhibition of the receptor could potentially counteract the disease evolution. The S1P₃ receptor appears to be an appealing target also for other therapeutic areas, in which a potential healing role of S1P₃ antagonism has been demonstrated.

S1P₃ Antagonism in Peripheral Diseases

S1P₃ activity has been shown to be implicated in inflammation-associated diseases such as arthritis (Lai et al., 2010), and several type of fibrosis (Shea and Tager, 2012) like heart (Takuwa et al., 2010), pulmonary (Kono et al., 2007), muscular (Cencetti et al., 2008) and liver fibrosis (Li et al., 2009) or in other more general inflammatory syndromes (Niessen et al., 2008) where S1P₃ receptor antagonism could potentially limit the pathologic processes.

S1P₃ receptor activation in the cardiovascular system could exert several pathologically-relevant effects. In the blood the S1P released by activated platelets stimulates S1P₃ (and S1P₁) receptors in vascular endothelium decreasing vascular para-cellular permeability (Mehta et al. 2005; Sun et al. 2009). Additionally, S1P₃ transactivation has been shown to disrupt vascular barrier regulation (Singleton et al., 2006). Furthermore, also the chemotactic effect of S1P in macrophages (demonstrated in vitro and in vivo) is mediated by S1P₃, so playing a causal role in atherosclerosis by promoting the recruitment of inflammatory monocyte/macrophage and altering vessel smooth muscle cells behaviour (Keul et al., 2011). Finally, the group of Takakura has demonstrated by a specific antagonist that the S1P-induced coronary flow decrease is dependent on S1P₃ receptor and so such antagonism might be adapt to counteract S1P related vascular diseases and vasospasm syndromes (Murakami et al., 2010). In the heart, interestingly, the sustained bradycardia induced by S1P receptor non-selective agonists is abolished in S1P₃ knockout mice or after S1P₃ pharmacological inhibition in rats (Sanna et al., 2004; Murakami et al., 2010). More, in the cardiac vascular microcirculation cells in diabetes, it has been shown in vivo and in vitro that the agonist FTY720 exerts a functional antagonism by stimulating the translocation of S1P₃ from membrane to the nucleus. Arguably, the pharmacological modulation of S1P₃ receptors could be beneficial to alleviate cardiac microangiopathy in diabetes (Yin et al., 2012).

Bajwa et al. (2012) have demonstrated that S1P plays a pivotal role in kidney ischemia-reperfusion injury (IRI). S1P₃ receptor-deficient mice were protected from IRI. This protective effect was due, at least in part to differences between S1P3-deficient dendritic cells. It was then supposed that pharmacological treatment are able to limit S1P₃ activity or treatments with dendritic cells lacking the S1P₃ receptor could help against progression of IRI.

Also several physiological parameters of the respiratory system are affected by S1P₃ activity. It has been recently demonstrated that the SIP pathway activation induced a generalized airway hyperreactivity in vivo and in vitro and this is mediated by S1P₃ receptor. Then, the S1P₃ antagonism, besides or contextually to the abovementioned putative healing effects on lung fibrosis, could represent a new therapeutic strategy aimed at blocking the asthma-related airway hyperreactivity (Trifilieff and Fozard, 2012). S1P₃ has been also shown to be strictly involved in acute lung injury where it promotes chemotaxis and increased endothelial and epithelial permeability (Uhlig and Yang, 2013). In the publication of Chen et al., (2008) it is suggested that S1P acting through S1P₃, increasing calcium influx, and Rho kinase, activates cPLA(2)alpha and releases arachidonic acid in lung epithelial cells. Then, understanding this mechanism in epithelial cells may provide potential targets to control inflammatory processes in the lung.

S1P₃ receptors play an important role in other non-inflammatory diseases. In cancer, it has been shown that S1P₃ activation promotes breast cancer cells invasiveness (Kim et al., 2011) and this effect can be diminished by a specific antibody able to block the receptor (Harris et al., 2012). Similar results were obtained in thyroid cancer cells (Balthasar et al., 2006) and glioma cells (Young et al., 2007), where S1P₃ activation showed to enhance cell migration and invasion. Yamashita (2006) also demonstrated that S1P₃-mediated signals might be crucial in determining the metastatic response of gastric cancer cells to SIP.

In the eye, considering that SIP is constitutively present in the aqueous humor (Liliom et al., 1998), and, in addition, that the endothelial cells of the trabecular meshwork, which express S1P₁ and S1P₃ receptors (Mettu et al., 2004), respond to S1P stimulus increasing the outflow resistance, the S1P₃ receptors pharmacological inhibition represents a potential therapeutic strategy in healing pathologies involving high intraocular pressure such as ocular hypertension, glaucoma, glaucomatous retinopathy (Stamer et al., 2009).

S1P₃ Antagonism in PNS Diseases

The tissue injury inflammation is associated with an increased sensitivity to noxious stimuli, suggesting that there could be an important interaction between the activities of immune cells and the sensory neurons activated by noxious stimulation. A direct exposure of isolated sensory neurons to S1P (together with other inflammatory signals released by platelets or mast cells) increases their action potential firing through activation of ion channels (Zhang et al., 2006). In experimental conditions of isolated sensory neurons, the expression of S1P3 receptors is the highest in the panel of S1P receptors. In addition, the Kress's laboratory has demonstrated that S1P3 receptor was detected in all human and mouse dorsal root ganglia neurons and that S1P evokes significant nociception via G-protein-dependent activation of an excitatory chloride conductance (Camprubí-Robles et al., 2013). Considering that S1P-induced neuronal responses and spontaneous pain behavior in vivo were strongly reduced in S1P3-null mice, S1P3 receptors could represent important therapeutic targets for post-traumatic pain (Camprubí-Robles et al., 2013).

S1P3 Antagonism in CNS Diseases

In the CNS, neurons, astrocytes, oligodendrocytes and microglia cells have the capacity to produce and secrete S1P and express, with different extents depending on the cell type, S1P₁₋₃ and S1P₅ receptors (Anelli et al., 2005; Foster et al., 2007). In regard to S1P₃ receptor, an intrinsic high expression has been seen in both astrocytes and neurons (Foster et al., 2007). S1P₃ is described to induce glial activation under pro-inflammatory conditions (Fisher et al., 2011; Wu et al., 2008) and enhance spontaneous glutamate release in the hippocampus mossy fibers (Kanno and Nishizaki, 2011). In particular, apoptotic neurons self-induce an overexpression of sphingosine-kinase with a further release of S1P. This process, elegantly demonstrated by Gude (Gude et al., 2008) and defined as “come-and-get-me” signal, has the purpose of chemo-attract microglial cells and eliminate the dying neuron. Furthermore, S1P through a G12/13 protein, by remodelling of actin cytoskeleton, can inhibit astrocytes tight junction, conferring them mobility, and creating gaps through the brain tissue (Rouach et al., 2006). Then, the action of S1P to astrocytes could help the activated microglial cells to better move in the brain and so express their phagocytic role. The S1P₃ receptors coupling to the G12/13 protein, associated to a high S1P₃ receptor expression in astrocytes and its role in motility (Fischer et al., 2011) leaded to the consideration that the described process could be conducted by a S1P₃-activated signalling. Interestingly, microglial cells, once activated, enhance their S1P₃ expression (Foster et al., 2007). With these evidences it was conceivably hypothesised that activation of S1P₃ receptor system is strictly involved in a neuroinflammatory state and S1P₃ inhibition could have limited its development. Evidence supporting S1P₃ antagonism to be protective in neuroinflammation was given from a mouse model of Sandhoff disease in which the ablation of the gene encoding S1P₃ receptor strongly limited the astroglial proliferation, prolonging the survival and improving motor function of the mice (Wu et al., 2008).

In neurodegenerative diseases neuroinflammation can play a clear detrimental role during the pathologic evolution Alzheimer's, Parkinson's, Amyotrophic lateral sclerosis, Huntington and Multiple sclerosis (Bradl and Hohlfeld, 2003; Maragakis and Rothstein, 2006; Davies et al., 2013). In Alzheimer disease (AD) the accumulation of beta-amyloid plaques has been associated to inflammation development with activation of the CNS immune system (Meraz-Rios et al., 2013). A relevance of S1P receptors activity and modulation in AD is also shown in Takasugi et al., 2011 and in Takasugi et al., 2013

PRIOR ART

EP81756 discloses compounds that are useful for treating inflammation.

Wang et al. (Bioorganic & Medicinal Chemistry Letters (2010), 20(2), 493-498) disclose compounds that are FFA2 inhibitors useful for the treatment of diabetes.

WO2000026202 discloses antiproliferative compounds that are useful for the treatment of cancer.

WO2003063797 discloses potassium channel inhibitors that are useful for the treatment of arrhythmia.

JP2002155065 discloses compounds that are useful as insecticides or miticides.

WO2001036415 discloses compounds that are useful for controlling pests on domestic animals and livestock.

In WO2005075435, WO2007087066, WO2008141119, WO2008147797, WO2009006315, WO2009123896 and WO2010054138, Vertex discloses compounds as CFTR modulators for the treatment of cystic fibrosis or as intermediates towards such compounds.

Ingyong Huaxue (1990), 7(6), 54-7 discloses pesticides and fungicides.

The following disclose activators of glucokinase useful for the treatment of diabetes: Journal of Medicinal Chemistry (2012), 55(3), 1318-1333; WO2009140624; US-20100063063; Journal of Medicinal Chemistry (2012), 55(16), 7021-7036; Medicinal Chemistry Letters (2013), 4(4), 414-418; US-20010039344; WO2001083465; WO2003095438; WO2004052869; WO2006058923; WO2007026761; WO2007041365; WO2008005914; WO2008078674; WO2008119734; WO2009091014; U.S. Pat. No. 6,610,846; Journal of Medicinal Chemistry (2010), 53(9), 3618-3625; WO2002046173; US20070281942; US2010063063.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1: Neuroprotective effect of a compound of the invention on quisqualic acid lesioned rats. Example 33 shows strong neuroprotective effects at 30 and 60 mg/Kg/day. Rat NBM ChAT-positive neuron counts using APERIO® (left), example 33 at doses of 30 and 60 mg/kg was administered once a day p.o.; the dose of 10 mg/kg was administered p.o. twice a day. In the microphotographs (right panel) a ChAT-qualitative analysis is shown in Quisqualic acid (QUIS) or Sham (SHAM)-injected rat NBM treated with vehicle or example 33 at 30 mg/kg p.o. *p<0.05 vs SHAM (Dunnet Test).

FIG. 2: Anti-neuroinflammatory effect of a compound on quisqualic acid lesioned rats. Example 33 has a strong activity on GFAP reactivity at 30 and 60 mg/Kg/day. A) effect on microglia cells, OX42 analysis; B) effect on astrocytes, GFAP analysis. These evidences are in line with the literature, in which the expression of S1P3 on microglia is considered low and not relevant (S1P2 is predominant), while the receptor appears to be highly expressed in astrocytes.

FIG. 3: Effect of a compound of the invention on Abeta(25-35) lesioned rats. Microscope scanning (GFAP staining) A) Ab (23-35) (right hemisphere)+Vehicle B) Ab(25-35) (right hemisphere)−example 33 at 30 mg/kg (left hemisphere was Sham-treated). C) quantitative analysis of GFAP-positive cells by visual scoring (*p<0.05 vs vehicle group, Kruskall-Wallis).

FIG. 4: Effect of a compound of the invention in the Object Recognition Test on quisqualic acid lesioned rats. Treatment with example 33 significantly ameliorates cognitive functions in ORT measuring episodic memory in Quisqualic lesioned rats as reported in the table of ANOVA (right panel). To note the difference in the exploration time between familiar and novel objects (F=familiar object, N=novel object).

DETAILED DESCRIPTION OF THE INVENTION

In a first aspect of this invention (embodiment A), there is provided compounds of formula (A),

wherein

●-R₁ is

X₁, X₆, X₇, X₉ and X₁₀ are halogen, C₁-C₄ linear branched or cyclic alkyl optionally substituted with one or more fluorine atoms; X₂, X₃, X₄, X₅ and X₈, are hydrogen, halogen, C₁-C₄ linear branched or cyclic alkyl optionally substituted with one or more fluorine atoms; with the proviso that at least one of X₂, X₃, X₄, and X₅ is not hydrogen; R₂ is a C₃-C₆ linear branched or cyclic alkyl optionally substituted with phenyl, with one or more fluorine atoms or with trifluoromethyl-furanyl; R₂′ is hydrogen, F, C₁-C₃ linear or branched alkyl optionally substituted with one or more fluorine atoms; or R₂ and R₂′ together with the carbon atom they are attached to form a C₃-C₆ cycloalkyl ring;

●-R₃ is

Y₁ is halogen; Y₁′ is C₁-C₃ linear branched or cyclic alkyl optionally substituted with one or more fluorine atoms; Y₂ is cyano or methoxyphenyl, C₁-C₃ linear branched or cyclic alkyl optionally substituted with one or more fluorine atoms; Y₃ is hydrogen, halogen or methoxyphenyl; Y₄ is hydrogen, halogen, N-methylpyrazolyl or a C₁-C₃ linear branched or cyclic alkoxy optionally substituted with one or more fluorine atoms, Y₅ is hydrogen, halogen, cyano, or a C₁-C₃ linear branched or cyclic alkyl optionally substituted with one or more fluorine atoms; with the proviso that at least one of Y₄ and Y₅ is not hydrogen; Y₆ is halogen, C₁-C₃ linear branched or cyclic alkyl optionally substituted with one or more fluorine atoms, or a C₁-C₃ linear branched or cyclic alkoxy optionally substituted with one or more fluorine atoms; enantiomers, enantiomerically enriched mixtures, and pharmaceutically acceptable salts thereof.

Under one aspect of embodiment A (embodiment A1), there is provided compounds of formula (A) wherein halogen is selected from the list of Cl, Br and F;

C₁-C₄ linear branched or cyclic alkyl optionally substituted with one or more fluorine atoms is selected from the list of methyl, trifluoromethyl, n-propyl and t-butyl; C₁-C₃ linear branched or cyclic alkyl optionally substituted with one or more fluorine atoms is selected from the list of methyl, trifluoromethyl and n-propyl; C₁-C₃ linear branched or cyclic alkoxy optionally substituted with one or more fluorine atoms is selected from the list of methoxy and difluoromethoxy; C₃-C₆ linear branched or cyclic alkyl optionally substituted with phenyl, with one or more fluorine atoms or with trifluoromethyl-furanyl is selected from n-propyl, 3-phenyl-n-propyl i-propyl, n-butyl, cyclohexyl and (5-trifluoromethyl-furan-2yl)-methyl; C₃-C₆ cycloalkyl is selected from the list of cyclobutyl and cyclopentyl;

Under another aspect of embodiment A (embodiment A2), there is provided compounds of formula (A) wherein halogen is selected from the list of Cl, Br and F;

C₁-C₄ linear branched or cyclic alkyl optionally substituted with one or more fluorine atoms is selected from the list of methyl, trifluoromethyl and t-butyl; C₁-C₃ linear branched or cyclic alkyl optionally substituted with one or more fluorine atoms is selected from the list methyl and trifluoromethyl; C₁-C₃ linear branched or cyclic alkoxy optionally substituted with one or more fluorine atoms is selected from the list of methoxy and difluoromethoxy; C₃-C₆ linear branched or cyclic alkyl optionally substituted with phenyl, with one or more fluorine atoms or with trifluoromethyl-furanyl is selected from n-propyl, 3-phenyl-n-propyl i-propyl, n-butyl, cyclohexyl and (5-trifluoromethyl-furan-2yl)-methyl; C₃-C₆ cycloalkyl is selected from list of cyclobutyl and cyclopentyl; Under another aspect of embodiment A (embodiment A3), there is provided compounds of formula (A) wherein R₁ and R₃ are as described under embodiment A and wherein X₁ is halogen X₂ is hydrogen, halogen or methyl X₃ is hydrogen, halogen or trifluoromethyl X₄ is hydrogen or methyl X₅ is hydrogen or halogen with the proviso that at least one of X₂, X₃, X₄, and X₅ is not hydrogen X₆ is halogen X₇ is t-butyl or trifluoromethyl, preferably t-butyl X₈ is hydrogen, methyl or t-butyl X₉ is halogen X₁₀ is t-butyl R₂ is n-propyl, 3-phenyl-n-propyl, i-propyl, n-butyl, cyclohexyl or (5-trifluoromethyl-furan-2yl)-methyl R₂′ is hydrogen, F, methyl or R₂ and R₂′ together with the carbon atom they are attached to form a cyclobutyl or cyclopentyl ring; Y₁ is halogen Y₁′ is methyl Y₂ is methyl, n-propyl, cyano, trifluoromethyl or 4-methoxyphenyl Y₃ is hydrogen, halogen, or 4-methoxyphenyl Y₄ is hydrogen, halogen, methoxy or 1-methyl-pyrazol-4-yl Y₅ is hydrogen, halogen, cyano or methyl with the proviso that at least one of Y₄ and Y₅ is not hydrogen Y₆ halogen, methoxy or difluoromethoxy

Under another aspect of embodiment A (embodiment A4), there is provided compounds of formula (A) wherein R₁ and R₃ are as described under embodiment A and

wherein

X₁ is Cl or Br

X₂ is hydrogen, methyl, Br or F X₃ is hydrogen, Br, Cl, F, or trifluoromethyl X₄ is hydrogen or methyl X₅ is hydrogen or F with the proviso that at least one of X₂, X₃, X₄, and X₅ is not hydrogen

X₆ is Cl

X₇ is t-butyl or trifluoromethyl, preferably t-butyl X₈ is hydrogen, methyl or t-butyl

X₉ is Br, Cl or F

X₁₀ is t-butyl R₂ is n-propyl, 3-phenyl-n-propyl, i-propyl, n-butyl, cyclohexyl or (5-trifluoromethyl-furan-2yl)-methyl; R₂′ is hydrogen, F, methyl; or R₂ and R₂′ together with the carbon atom they are attached to form a cyclobutyl or cyclopentyl ring;

Y₁ is Br

Y₁′ is methyl Y₂ is methyl, n-propyl, cyano, trifluoromethyl and 4-methoxyphenyl Y₃ is hydrogen, Br, Cl, and 4-methoxyphenyl Y₄ is hydrogen, Br, Cl, methoxy or 1-methyl-pyrazol-4-yl Y₅ is hydrogen, Br, Cl, F, cyano or methyl with the proviso that at least one of Y₄ and Y₅ is not hydrogen Y₆ is Br, Cl, F, methoxy or difluoromethoxy

Under specific aspects of embodiments A, A1, A2, A3 or A4 (embodiments B1) there is provided compounds of formula (A) wherein ●-R₁ is selected from

and wherein R₂, R₂′, R₃, X₁, X₂, X₃, X₄, X₅, X₆, X₇, X₈, X₉, X₁₀, Y₁, Y₁′, Y₂, Y₃, Y₄, Y₅ and Y₆ are, as the case may be, as defined under embodiment A, A1, A2, A3 or A4

Under specific aspects of embodiments A, A1, A2, A3 or A4 (embodiments B2) there is provided compounds of formula (A) wherein ●-R₁ is selected from

and wherein R₂, R₂′, R₃, X₇, Y₁, Y₁′, Y₂, Y₃, Y₄, Y₅ and Y₆ are, as the case may be, as defined under embodiment A, A1, A2, A3 or A4

Under specific aspects of embodiments A, A1, A2, A3 or A4 (embodiments B3) there is provided compounds of formula (A) wherein ●-R₁ is selected from

and wherein R₂, R₂′, R₃, X₁, Y₁, Y₁′, Y₂, Y₃, Y₄, Y₅ and Y₆ are, as the case may be, as defined under embodiment A, A1, A2, A3 or A4

Under specific aspects of embodiments A, A1, A2, A3 or A4 (embodiments B4) there is provided compounds of formula (A) wherein ●-R₁ is selected from

and wherein R₂, R₂′, R₃, X₂, X₃, X₄, X₅, Y₁, Y₁′, Y₂, Y₃, Y₄, Y₅ and Y₆ are, as the case may be, as defined under embodiment A, A1, A2, A3 or A4

Under a particular aspect of embodiments B4 (embodiments B4a), there is provided compounds of formula (A) wherein X₂ and X₄ are hydrogen and wherein R₁, R₂, R₂′, R₃, X₃, X₅, Y₁, Y₁′, Y₂, Y₃, Y₄, Y₅ and Y₆ are, as the case may be, as defined under embodiments B4.

Under another particular aspect of embodiments B4 (embodiments B4b), there is provided compounds of formula (A) wherein X₂, X₄ and X₅ are hydrogen and wherein R₁, R₂, R₂, R₃, X₃, Y₁, Y₁′, Y₂, Y₃, Y₄, Y₅ and Y₆ are, as the case may be, as defined under embodiments B4.

Under specific aspects of embodiments A, A1, A2, A3 or A4 (embodiments B5) there is provided compounds of formula (A) wherein ●-R₁ is selected from

and wherein R₂, R₂′, R₃, X₆, Y₁, Y₁′, Y₂, Y₃, Y₄, Y₅ and Y₆ are, as the case may be, as defined under embodiment A, A1, A2, A3 or A4

Under specific aspects of embodiments A, A1, A2, A3 or A4 (embodiments B6) there is provided compounds of formula (A1 wherein ●-R₁ is selected from

and wherein R₂, R₂′, R₃, X₇, Y₁, Y₁′, Y₂, Y₃, Y₄, Y₅ and Y₆ are, as the case may be, as defined under embodiment A, A1, A2, A3 or A4

Under specific aspects of embodiments A, A1, A2, A3 or A4 (embodiments B7) there is provided compounds of formula (A) wherein ●-R₁ is selected from

and wherein R₂, R₂′, R₃, X₈, X₉, Y₁, Y₁′, Y₂, Y₃, Y₄, Y₅ and Y₆ are, as the case may be, as defined under embodiment A, A1, A2, A3 or A4

Under specific aspects of embodiments A, A1, A2, A3 or A4 (embodiments B8) there is provided compounds of formula (A) wherein ●-R₁ is selected from

and wherein R₂, R₂′, R₃, X₁₀, Y₁, Y₁′, Y₂, Y₃, Y₄, Y₅ and Y₆ are, as the case may be, as defined under embodiment A, A1, A2, A3 or A4

Under specific aspects of embodiments A, A1, A2, A3, A4, B1, B2, B3, B4, B4a, B4b, B5, B6, B7 or B8 (embodiments C1) there is provided compounds of formula (A) wherein ●-R₃ is selected from

and wherein R₁, R₂, R₂′, X₁, X₂, X₃, X₄, X₅, X₆, X₇, X₈, X₉, X₁₀, Y₁, Y₂, and Y₃ are, as the case may be, as defined under embodiments A, A1, A2, A3, A4, B1, B2, B3, B4, B4a, B4b, B5, B6, B7 or B8

Under specific aspects of embodiments A, A1, A2, A3, A4, B1, B2, B3, B4, B4a, B4b, B5, B6, B7 or B8 (embodiments C2) there is provided compounds of formula (A) wherein ●-R₃ is selected from

and wherein R₁, R₂, R₂′, X₁, X₂, X₃, X₄, X₅, X₆, X₇, X₈, X₉, X₁₀, Y₁, Y₄, Y₅, and Y₆ are, as the case may be, as defined under embodiments A, A1, A2, A3, A4, B1, B2, B3, B4, B4a, B4b, B5, B6, B7 or B8

Under specific aspects of embodiments A, A1, A2, A3, A4, B1, B2, B3, B4, B4a, B4b, B5, B6, B7 or B8 (embodiments C3) there is provided compounds of formula (A) wherein ●-R₃ is selected from

and wherein R₁, R₂, R₂′, X₁, X₂, X₃, X₄, X₅, X₆, X₇, X₈, X₉, X₁₀, Y₁, Y₁′, Y₄, Y₅, and Y₆ are, as the case may be, as defined under embodiments A, A1, A2, A3, A4, B1, B2, B3, B4, B4a, B4b, B5, B6, B7 or B8

Under specific aspects of embodiments A, A1, A2, A3, A4, B1, B2, B3, B4, B4a, B4b, B5, B6, B7 or B8 (embodiments C4) there is provided compounds of formula (A) wherein ●-R₃ is selected from

and wherein R₁, R₂, R₂′, X₁, X₂, X₃, X₄, X₅, X₆, X₇, X₈, X₉, X₁₀ and Y₁ are, as the case may be, as defined under embodiments A, A1, A2, A3, A4, B1, B2, B3, B4, B4a, B4b, B5, B6, B7 or B8

Under specific aspects of embodiments A, A1, A2, A3, A4, B1, B2, B3, B4, B4a, B4b, B5, B6, B7 or B8 (embodiments C5) there is provided compounds of formula (A) wherein ●-R₃ is selected from

and wherein R₁, R₂, R₂′, X₁, X₂, X₃, X₄, X₅, X₆, X₇, X₈, X₉, X₁₀ and Y₁ are, as the case may be, as defined under embodiments A, A1, A2, A3, A4, B1, B2, B3, B4, B4a, B4b, B5, B6, B7 or B8

Under specific aspects of embodiments A, A1, A2, A3, A4, B1, B2, B3, B4, B4a, B4b, B5, B6, B7 or B8 (embodiments C6) there is provided compounds of formula (A) wherein ●-R₃ is selected from

and wherein R₁, R₂, R₂′, X₁, X₂, X₃, X₄, X₅, X₆, X₇, X₈, X₉, X₁₀, Y₂ and Y₃ are, as the case may be, as defined under embodiments A, A1, A2, A3, A4, B1, B2, B3, B4, B4a, B4b, B5, B6, B7 or B8

Under specific aspects of embodiments A, A1, A2, A3, A4, B1, B2, B3, B4, B4a, B4b, B5, B6, B7 or B8 (embodiments C7) there is provided compounds of formula (A) wherein ●-R₃ is selected from

and wherein R₁, R₂, R₂′, X₁, X₂, X₃, X₄, X₅, X₆, X₇, X₈, X₉, X₁₀ and Y₁ are, as the case may be, as defined under embodiments A, A1, A2, A3, A4, B1, B2, B3, B4, B4a, B4b, B5, B6, B7 or B8

Under specific aspects of embodiments A, A1, A2, A3, A4, B1, B2, B3, B4, B4a, B4b, B5, B6, B7 or B8 (embodiments C8) there is provided compounds of formula (A) wherein ●-R₃ is selected from

and wherein R₁, R₂, R₂′, X₁, X₂, X₃, X₄, X₅, X₆, X₇, X₈, X₉, X₁₀ and Y₄ and Y₅ are, as the case may be, as defined under embodiments A, A1, A2, A3, A4, B1, B2, B3, B4, B4a, B4b, B5, B6, B7 or B8

Under specific aspects of embodiments A, A1, A2, A3, A4, B1, B2, B3, B4, B4a, B4b, B5, B6, B7 or B8 (embodiments C9) there is provided compounds of formula (A) wherein ●-R₃ is selected from

and wherein R₁, R₂, R₂′, X₁, X₂, X₃, X₄, X₅, X₆, X₇, X₈, X₉, X₁₀ and Y₆ are, as the case may be, as defined under embodiments A, A1, A2, A3, A4, B1, B2, B3, B4, B4a, B4b, B5, B6, B7 or B8

Under specific aspects of embodiments A, A1, A2, A3, A4, B1, B2, B3, B4, B4a, B4b, B5, B6, B7 or B8 (embodiments C10) there is provided compounds of formula (A) wherein ●-R₃ is selected from

and wherein R₁, R₂, R₂′, X₁, X₂, X₃, X₄, X₅, X₆, X₇, X₈, X₉ and X₁₀ are, as the case may be, as defined under embodiments A, A1, A2, A3, A4, B1, B2, B3, B4, B4a, B4b, B5, B6, B7 or B8.

Under specific aspects of embodiments A, A1, A2, A3, A4, B1, B2, B3, B4, B4a, B4b, B5, B6, B7 and B8 (embodiments C11) there is provided compounds of formula (A) wherein ●-R₃ is selected from

and wherein R₁, R₂, R_(2′), X₁, X₂, X₃, X₄, X₅, X₆, X₇, X₈, X₉ and X₁₀ are, as the case may be, as defined under embodiments A, A1, A2, A3, A4, B1, B2, B3, B4, B4a, B4b, B5, B6, B7 or B8.

Under specific aspects of embodiments A, A1, A2, A3, A4, B1, B2, B3, B4, B4a, B4b, B5, B6, B7, B8, C1, C2, C3, C4, C5, C6, C7, C8, C9, C10 and C11 (embodiments D1) there is provided compounds of formula (A) wherein R₂ and R₂′ do not form a cycloalkyl ring together with the carbon atom they are attached to.

and wherein R₁, R₃, X₁, X₂, X₃, X₄, X₅, X₆, X₇, X₈, X₉, X₁₀, Y₁, Y₁′, Y₂, Y₃, Y₄, Y₅ and Y₆ are, as the case may be, as defined under embodiments A, A1, A2, A3, A4, B1, B2, B3, B4, B4a, B4b, B5, B6, B7, B8, C1, C2, C3, C4, C5, C6, C7, C8, C9, C10 or C11.

Under particular aspects of embodiments D1 (embodiments D1a), there is provided compounds of formula (A) wherein R₂′ is hydrogen and wherein R₁, R₂, R₃, X₁, X₂, X₃, X₄, X₅, X₆, X₇, X₈, X₉, X₁₀, Y₁, Y₁′, Y₂, Y₃, Y₄, Y₅ and Y₆ are, as the case may be, as defined under embodiments D1.

Under other particular aspects of embodiments D1 (embodiments D1b), there is provided compounds of formula (A) wherein R₂′ is F and wherein R₁, R₂, R₃, X₁, X₂, X₃, X₄, X₅, X₆, X₇, X₈, X₉, X₁₀, Y₁, Y₁′, Y₂, Y₃, Y₄, Y₅ and Y₆ are, as the case may be, as defined under embodiments D1.

Under another particular aspect of embodiments D1 (embodiments D1c), there is provided compounds of formula (A) wherein R₂′ is different from hydrogen or F and wherein R₁, R₂, R₃, X₁, X₂, X₃, X₄, X₅, X₆, X₇, X₈, X₉, X₁₀, Y₁, Y₁′, Y₂, Y₃, Y₄, Y₅ and Y₆ are, as the case may be, as defined under embodiments D.

Under specific aspect of embodiments D1a, D1b and D1c (embodiments D1d), there is provided compounds of formula (A) wherein R₂ is n-propyl and wherein R₁, R₂′, R₃, X₁, X₂, X₃, X₄, X₅, X₆, X₇, X₈, X₉, X₁₀, Y₁, Y₁′, Y₂, Y₃, Y₄, Y₅ and Y₆ are, as the case may be, as defined under embodiments D1a, D1b and D1c.

Under specific aspect of embodiments D1a, D1b and D1c (embodiments D1e), there is provided compounds of formula (A) wherein R₂ is i-propyl and wherein R₁, R₂′, R₃, X₁, X₂, X₃, X₄, X₅, X₆, X₇, X₈, X₉, X₁₀, Y₁, Y₁′, Y₂, Y₃, Y₄, Y₅ and Y₆ are, as the case may be, as defined under embodiments D1a, D1b and D1c.

Under specific aspect of embodiments D1a, D1b and D1c (embodiments D1f), there is provided compounds of formula (A) wherein R₂ is n-butyl and wherein R₁, R₂′, R₃, X₁, X₂, X₃, X₄, X₅, X₆, X₇, X₈, X₉, X₁₀, Y₁, Y₁′, Y₂, Y₃, Y₄, Y₅ and Y₆ are, as the case may be, as defined under embodiments D1a, D1b and D1c.

Under specific aspect of embodiments D1a, D1b and D1c (embodiments D1f), there is provided compounds of formula (A) wherein R₂ is cyclohexyl and wherein R₁, R₂′, R₃, X₁, X₂, X₃, X₄, X₅, X₆, X₇, X₈, X₉, X₁₀, Y₁, Y₁′, Y₂, Y₃, Y₄, Y₅ and Y₆ are, as the case may be, as defined under embodiments D1a, D1b and D1c.

Under specific aspects of embodiments A, A1, A2, A3, A4, B1, B2, B3, B4, B4a, B4b, B5, B6, B7, B8, C1, C2, C3, C4, C5, C6, C7, C8, C9, C10 and C11 (embodiments D2) there is provided compounds of formula (A) wherein R₂ and R₂′ together with the carbon atom they are attached to form a cycloalkyl ring and wherein R₁, R₃, X₁, X₂, X₃, X₄, X₅, X₆, X₇, X₈, X₉, X₁₀, Y₁, Y₁′, Y₂, Y₃, Y₄, Y₅ and Y₆ are, as the case may be, as defined under embodiments A, A1, A2, A3, A4, B1, B2, B3, B4, B4a, B4b, B5, B6, B7, B8, C1, C2, C3, C4, C5, C6, C7, C8, C9, C10 and C11.

The combination of any of the above embodiments gives rise to a new embodiment under this invention.

For example, from the combination of embodiments B3, B4 and B7 there is provided other specific aspects of embodiments A, A1, A2, A3 or A4 (Embodiments E1) which are compounds of formula (A) wherein ●-R₁ is selected from

and wherein R₂, R₂′, R₃, X₁, X₂, X₃, X₄, X₅, X₈, X₉, Y₁, Y₁′, Y₂, Y₃, Y₄, Y₅ and Y₆ are, as the case may be, as defined under embodiment A, A1, A2, A3 or A4

Likewise, from the combination of embodiments B3, B4a and B7, there is provided other specific aspects of embodiments A, A1, A2, A3 or A4 (Embodiments E1a) which are compounds of formula (A) wherein ●-R₁ is selected from

X₂ and X₄ are hydrogen;

and wherein R₂, R₂′, R₃, X₁, X₃, X₅, X₈, X₉, Y₁, Y₁′, Y₂, Y₃, Y₄, Y₅ and Y₆ are, as the case may be, as defined under embodiment A, A1, A2, A3 or A4

Also, from the combination of embodiments C6, C8, C9 and C11 there is provided other specific aspects of embodiments A, A1, A2, A3, A4, B1, B2, B3, B4, B4a, B4b, B5, B6, B7 or B8 (Embodiments E2) which are compounds of formula (A) wherein ●-R₃ is selected from

and wherein R₁, R₂, R₂′, X₁, X₂, X₃, X₄, X₅, X₆, X₇, X₈, X₉, X₁₀, Y₂, Y₃, Y₄, Y₅ and Y₆ are, as the case may be, as defined under embodiments A, A1, A2, A3, A4, B1, B2, B3, B4, B4a, B4b, B5, B6, B7 or B8.

Examples 1-151 described below all constitute further individual embodiments of this invention, and any list combining any of the examples is yet another further embodiment of this invention.

For example, in a further embodiment (embodiment F1) there is provided a compound selected form the list of

-   1-(5-Chloro-pyridin-3-yl)-cyclobutanecarboxylic acid     (5-chloro-pyridin-2-yl)-amide; -   2-(6-Chloro-5-cyano-pyridin-3-yl)-pentanoic acid     (5-chloro-pyrazin-2-yl)-amide; -   2-(6-Chloro-5-fluoro-pyridin-3-yl)-pentanoic acid     (5-bromo-pyrazin-2-yl)-amide; -   2-(6-Bromo-pyridin-2-yl)-pentanoic acid     (5-bromo-pyrazin-2-yl)-amide; -   2-(2-Bromo-pyridin-4-yl)-pentanoic acid     (5-bromo-pyridin-2-yl)-amide; -   2-(2-Methoxy-pyridin-4-yl)-pentanoic acid     (5-bromo-pyridin-2-yl)-amide; -   2-(6-Methoxy-pyridin-3-yl)-pentanoic acid     (5-bromo-pyridin-2-yl)-amide; -   2-(4-Bromo-3-trifluoromethyl-pyrazol-1-yl)-pentanoic acid     (5-bromo-pyrazin-2-yl)-amide; -   2-(2-Difluoromethoxy-pyridin-4-yl)-pentanoic acid     (5-bromo-pyridin-2-yl)-amide; -   2-[6-(1-Methyl-1H-pyrazol-4-yl)-pyridin-3-yl]-pentanoic acid     (5-chloro-thiazol-2-yl)-amide; -   2-(5-Bromo-pyridin-3-yl)-pentanoic acid     (5-bromo-pyrazin-2-yl)-amide; -   2-(5-Bromo-pyridin-3-yl)-pentanoic acid     (5-bromo-3-methyl-pyridin-2-yl)-amide; -   2-(5-Bromo-pyridin-3-yl)-pentanoic acid     (5-bromo-6-fluoro-pyridin-2-yl)-amide; -   2-(5-Bromo-pyridin-3-yl)-pentanoic acid     (5-chloro-pyrazin-2-yl)-amide; -   2-(5-Bromo-pyridin-3-yl)-2-fluoro-pentanoic acid     (5-chloro-pyridin-2-yl)-amide; -   2-(2-Bromo-pyridin-4-yl)-pentanoic acid     (5-bromo-pyrazin-2-yl)-amide; -   2-(2-Bromo-pyridin-4-yl)-pentanoic acid     (5-chloro-pyridin-2-yl)-amide; -   2-(5-Bromo-pyridin-3-yl)-pentanoic acid     (5-fluoro-pyridin-2-yl)-amide; -   2-(2-Methoxy-pyridin-4-yl)-pentanoic acid     (5-bromo-thiazol-2-yl)-amide; -   2-(2-Methoxy-pyridin-4-yl)-pentanoic acid     (5-bromo-pyrazin-2-yl)-amide; -   2-(4-Bromo-3-trifluoromethyl-pyrazol-1-yl)-pentanoic acid     (5-bromo-pyridin-2-yl)-amide; -   2-(4-Bromo-3-cyano-pyrazol-1-yl)-pentanoic acid     (5-bromo-pyridin-2-yl)-amide; -   2-(5-Bromo-pyridin-3-yl)-hexanoic acid (5-bromo-pyrazin-2-yl)-amide; -   2-(4-[4-methoxy-phenyl]-3-trifluoromethyl-pyrazol-1-yl)-pentanoic     acid (5-bromo-pyrazin-2-yl)-amide; -   2-(4-Bromo-3-methyl-pyrazol-1-yl)-pentanoic acid     (5-bromo-pyrazin-2-yl)-amide; -   2-(4-Bromo-imidazol-1-yl)-pentanoic acid     (5-bromo-pyridin-2-yl)-amide; -   2-[3-(4-Methoxy-phenyl)-pyrazol-1-yl]-pentanoic acid     (5-bromo-pyrazin-2-yl)-amide; -   2-(4-Bromo-3-cyano-pyrazol-1-yl)-pentanoic acid     (5-bromo-pyrazin-2-yl)-amide; -   2-[5-Fluoro-6-(1-methyl-1H-pyrazol-4-yl)-pyridin-3-yl]-pentanoic     acid (5-chloro-pyrazin-2-yl)-amide; -   2-[5-Cyano-6-(1-methyl-1H-pyrazol-4-yl)-pyridin-3-yl]-pentanoic acid     (5-chloro-pyrazin-2-yl)-amide; -   2-(5-Bromo-pyridin-3-yl)-N-(5-bromo-pyrazin-2-yl)3-methyl-butyramide; -   N-(5-Bromo-3-fluoro-pyridin-2-yl)-2-(5-bromo-pyridin-3-yl)-3-methyl-butyramide; -   N-(5-Bromo-pyrazin-2-yl)-2,2-dicyclohexyl-acetamide; -   1-(5-Bromo-pyridin-3-yl)-cyclobutanecarboxylic acid     (5-bromo-pyrazin-2-yl)-amide; -   1-(5-Chloro-pyridin-3-yl)-cyclobutanecarboxylic acid     (5-bromo-pyrazin-2-yl)-amide; -   2-(6-Chloro-5-cyano-pyridin-3-yl)-pentanoic acid     (5-bromo-pyrazin-2-yl)-amide; -   2-(5-Chloro-pyridin-3-yl)-pentanoic acid     (5-bromo-pyrazin-2-yl)amide; -   2-(5-Chloro-pyridin-3-yl)-pentanoic acid     (5-chloro-pyrazin-2-yl)-amide; -   2-(6-Chloro-5-methyl-pyridin-3-yl)-pentanoic acid     (5-chloro-pyrazin-2-yl)-amide; -   and 2-(2-Chloro-pyridin-4-yl)-pentanoic acid     (5-bromo-pyrazin-2-yl)-amide

Likewise, in a in a further embodiment (embodiment F2) there is provided a compound selected form the list of 2-(5-Bromo-pyridin-3-yl)-pentanoic acid (3-tert-butyl-isoxazol-5-yl)-amide and 2-(5-Bromo-pyridin-3-yl)-hexanoic acid (3-tert-butyl-isoxazol-5-yl)-amide

General Route to Compounds of the Invention

Depending on the exact nature of the compound, compounds of the invention may be obtained under general Schemes 1-5.

Compounds with general structure I can be prepared as shown in Scheme 1. The key step of the synthesis is coupling between acids of general structure II and the appropriate amines of general structure III using coupling agents known in literature. Alternatively, amides of structure I can be achieved directly from esters IV where R2 is alkyl and R2′ can be fluorine or hydrogen. When acids of general structure II are not commercially available, they can be prepared according to different approaches.

Alkylation of commercially available heteroarylacetic acid of general structure V, with the appropriate haloalkane, in presence of a strong bases such LiHMDS, n-butyllithium, sodium hydride and other known in literature gives acids of general structure II.

When R2 and R2′ are different alkyl groups, acids of general structure II can be prepared in two steps. Heteroaryl acetic acids of general structure V can be alkylated to give intermediates of general structure VI, which undergo to a second alkylation to furnish acids of general structure II.

Alternatively commercially available heteroarylacetonitriles of general structure VII can be alkylated affording intermediates of general structure VIII which can be hydrolyzed to acids II.

Acids of general structure II can be obtained from hydrolysis of esters of general structure IV as reported in Scheme 1 where Pg can be methyl, ethyl or tert-butyl group. In case R2′ is a cyano group, hydrolysis and mono-decarboxylation occur simultaneously.

When esters of general structure IV are not commercially available they can be prepared following different synthetic pathways shown in Scheme 1.

Esters of general structure IV where R2 is an alkyl group and R2′ is fluorine can be prepared from acids VI, which are converted to corresponding esters IX and finally fluorinated in presence of a strong base and an electrophilic source of fluorine as reported by Tengeiji et al. Molecules 2012, 17, 7356-7378.

Esters IV can be prepared from heteroaryl bromides of structure X which are transformed into intermediates XI via palladium catalyzed reaction with malonitrile for example see Xiang Wang et al. J. Org. Chem., 2008, 73, 1643-1645. Derivatives XI can be alkylated to give esters IV where R2′ is cyano group.

Ester IV can be prepared directly from alkylation of heteroaryl acetic esters of general structure XII.

When esters XII are not commercially available, they can be synthesised with three different approaches: by alcoholysis of heteroaryl acetonitriles of general structure VII, via Negishi-Reformatsky coupling between heteroaryl bromides of structure X and tert-butyl ester of bromoacetic acid as described by Hartwig, J. F. et al. JACS, 2003, 125, 11176-11177 or by carboxylation of methyl group of compounds of general structure XIII in presence of a strong base as LDA (see WO9815278).

Esters of general structure IV can be prepared from N-alkylation of nitrogen bearing heterocycles XIV (pyrazoles, pyrroles, indoles) with ester of α-bromoalkanoic acid of general structure XV.

An alternative approach for the synthesis of compounds of general structure I, depicted in Scheme 1, consists of coupling between appropriate amine of general structure III with α-bromoacid of general structure XVI using a suitable coupling agent to give intermediate of general structure XVII. N-alkylation of nitrogen bearing heterocycles XIV (pyrazoles, pyrroles, and indoles) with intermediates XVII offer compounds of general structure I.

Compounds of general structure I can be further modified into derivatives Ia when R3 contains groups that can be modified in few synthetic steps. For example, when R3 contains methoxy group, it can be demethylated and O-alkylated with different alkyl groups; or in presence of primary amino group, this one can be alkylated to the corresponding tertiary amine or can be acylated with the appropriate carboxylic acid.

Scheme 2 describes the synthetic approach to prepare compounds of general structure Ib, where R3 is 1,2,4-oxadiazole substituted in position 3 with an aryl or heteroaryl group. Condensation between diethyl alkyl malonate XVIII and amidoxime XIX gives oxadiazole XX, which can be hydrolyzed to corresponding acid XXI and coupled with the appropriate amine of general structure III using a suitable coupling agent to give compounds of general structure Ib.

In Scheme 4 it is depicted the synthesis for a single point modification of intermediates of general structure IV, where methoxy group is replaced by difluoromethoxy moiety. Methoxypyridine esters of general structure IV are converted into the corresponding pyridones XXII followed by difluoromethylation of the oxygen (Makoto et al. Organic Letters, 2006, 8, 3805-3808) to give esters of general structure IVa. Direct coupling with the appropriate amines of general structure III affords final compounds I.

Scheme 4 depicts possible approaches for the synthesis of compounds of general structure I, where R3 is a bis-heteroaryl system. These syntheses can be applied on intermediates of general structure XII, V and VII containing a halogen in the R3 system. Intermediates of general structure II, IV, VIII can be obtained respectively from compounds V, XII and VII as described in Scheme 1. Suzuki coupling on II, IV and VIII gives compounds of general structure IIb, IVb and VIIIb. Intermediate IVb and VIIIb can be hydrolyzed to give compounds of general structure IIb which are converted into compounds I reacting with the appropriate amines of general structure III, using a suitable coupling agent.

Enantiomers or enantiomerically enriched compositions could also be obtained by using optically active starting materials or by enantiomeric resolution strategies.

Enantiomeric resolution strategies of compounds of general structure I are reported in scheme 5. Racemic mixtures of compounds I can be separated by chiral preparative HPLC.

Alternatively, acid of general structure II can be coupled with chiral auxiliaries such oxazolidinones to give diastereoisomers XXIIIa and XXIIIb. The resulting diastereoisomers could be separated and hydrolyzed to give the two acids in pure enantiomeric form that could be coupled with the appropriate amine of general structure III, using a suitable coupling agent to give compounds of general structure I as enantiomers. Alternatively racemic acids of general structure II can be solved before amide coupling with conventional approaches such as crystallization in presence of a chiral amine, enzymatic resolution, chiral preparative HPLC.

Enantiomers or enantiomerically enriched compositions could also be obtained by using optically active starting materials.

Biological Evaluation

In Vitro Cellular Assay for Activity Against S1P

CHO-S1P3 R1 cells were generated by stably transfecting wt CHO-K1 cells with pcDNA6.2/cLumioDEST-hS1P3 and were maintained under antibiotic selection with 6 μg/ml blasticidin. CHO-S1P1 MG12 cells were also generated by stable transfection of wt CHO-K1 cells and were selected with 1 mg/ml hygromycin.

The compounds were tested on CHO-S1P3 R1 cells for their ability to act as antagonists of the sphyngosine induced intracellular Ca-flux, that was measured by the fluorescent calcium indicator Fluo-4 AM on a Molecular Devices FLIPR3 instrument.

Cells were seeded as 30K cells per well in a 96-well plate (black, clear bottom, TC coated) in 100 μl culture medium. After 24 h incubation cells were loaded with 100 μL HBSS containing 20 mM HEPES buffer, 5 mM probenecid, 4 μM FLUO-4 AM and pluronic acid 0.02% and kept at 37° C., 5% CO2 for 30 minutes. Loading solution was then washed out with HBSS— 20 mM Hepes buffer.

Compounds were dispensed to the cells as first addition, at the final concentration of 10 μM for primary screening and in 8 points concentration response (30 μM-0.001 μM) with a final DMSO concentration of 0.3%. Sphingosine was added as second addition at the final concentration equal to the EC80. Calcium responses were read on a fluorescence imaging plate reader (FLIPR3; Molecular Devices) by exciting the cells with an argon ion laser at 488 nm. Emission was recorded by using a band spectrum filter (510-570 nm; emission peak of Fluo-4/Ca2+=516 nm).

The compound activity was also evaluated for the activity on Gai pathway both against S1P1 receptor and S1P3 receptor.

Changes in intracellular cAMP concentrations were measured with a HTRF® assay (cAMP Dynamic 2 Kit, Cisbio Bioassays, Codolet, France), according to the manufacturer's protocol.

CHO-S1P1 MG12 or CHO-S1P3 R1 ready-to-use frozen cells were thawed, resuspended in DPBS (Lonza, Basel, Switzerland) with 1 mM IBMX and dispensed in 384-well low volume microplates (Greiner Bio-One GmbH, Frickenhausen, Germany) as 10000 cells in 5 μl per well. Cell treatment was performed in assay buffer containing PBS 0.2% BSA. The cells were pre-incubated for 15 min at room temperature with 2.5 μl of a 4-fold concentrated compound solution either at single concentration or in a concentration response titration (0.6% final DMSO concentration). Subsequently, 2.5 μl of a Sphingosine/forskolin solution at 4-fold the respective EC80 concentrations were added to each well except for positive control wells, where only forskolin was added. After 45 min, 5 μl of the HTRF® detection reagents (anti cAMP-Cryptate and cAMP-d2) were added to the cells according to the kit instructions. After 1 hour incubation at room temperature, the time resolved fluorescence was read with an AnalystGT microplate reader (Molecular Devices, Sunnyvale, Calif., USA) with excitation at 337 nm, emission at 665 nm and 620 nm (for acceptor and donor signals, respectively).

In Vitro Phenotypic Assay: S1P Proliferation Assay in Primary Cortical Astrocytes

Primary cortical astrocytes were prepared from E17 embryos (Sprague-Dawley) rat neocortex by enzymatic dissociation. The isolated cortices were minced into small pieces with a sterile blade, washed for three times with dissociation medium and incubated with trypsin (0.25%) in waterbath at 37° C. for 10 mins. The pellet was placed in FBS (10%)-containing MEM medium and pipetted for 20 strokes; the cell suspension was centrifuged at 1050 rpm for 10 mins at r.t. and the pellet was resuspended in growth medium (BME, 10% FBS, 2 mM glutamine, 1 mM pyruvate, Penicillin/Streptomycin 1000 U/ml). The cells were seeded in poly-D-lysine (70K-150K kD) pre-coated 75 cm² flasks on at, at 37° C., 5% CO₂ and 95% humidity. The mature cultures were grown until astrocytes had reached confluence (12-15 days). Then, cultures are placed in an orbital shaker and shacked (200 RPM) overnight at 37° C., 5% CO₂ and 95% humidity. Medium was then removed and the cell layer containing mostly astrocytes was removed by trypsinization (0.25%) for 15 mins at 37° C. Furthermore, after blocking trypsin with MEM 10% FBS, medium was eliminated by centrifugation at 1200 RPM. Cells are seeded into black wall, clear bottom 96-well plates (30K cells/well) in 10% FBS/BME (day 1). 24 h later, on day 2, the medium was replaced with serum free BME. On day 3, the cells were treated with the antagonists for 1 hr before S1P addition (S1P final concentration: 1 μM). The final DMSO concentration was 0.1% v/v. On day 5 (48 h S1P stimulation), cells were fixed in 4% paraformaldehyde/4% sucrose, permeabilized in 0.2% Triton-X 100, and blocked in 0.1% BSA. The primary antibody Rb-anti-Ki67 (1:500, Abcam) was incubated for 3 hr at room temperature, followed by the Alexa Fluor 546 conjugated secondary antibody. The plates were acquired with BD Pathway 435 and the nuclear intensity of the Ki67 staining measured with BD Attovision software. The proliferation was expressed as percentage of Ki67-positive nuclei per total nuclei.

Neurodegeneration, Neuroinflammation and Behavioural In Vivo Assays.

The efficacy of the compounds of the invention on neurodegeneration and neuroinflammation can be evaluated with two different methodological approaches aimed to reproduce some pathological features of Alzheimer's disease:

-   1) the excitotoxic insult (quisqualic acid—QUIS) in the Nucleus     Basalis Magnocellularis (NBM), characterized by severe     neurodegeneration of cholinergic neurons along with significant     neuroinflammation. -   2) the β-amyloid peptide 25-35 (Aβ25-35) injection in NBM of rats,     inducing a significant glia reaction around Aβ25-35 deposits with     modest toxicity on cholinergic neurons.     -   Readouts are based on immunochemical analysis and are: count of         cholinergic neurons (ChAT-positive), astrocytes (GFAP-positive)         and microglia (OX-42 or Iba-1 positive). The analysis is         performed by two different approaches, the visual scoring         (blind) and the digital platform APERIO®.

QUIS-treated animals can also undergo the Object Recognition Test (ORT, measuring episodic memory) or other behavioural tests to measure the improvement of cognitive functions upon treatment with a compound of the invention.

Animals

Three-month old male Wistar rats (Harlan, Milan, Italy) weighing 230-250 g were used. The rats were housed in macrolon cages with ad lib food and water and maintained on a 12 h light/dark cycle at 23° C. room temperature (RT). All experiments were carried out according to the guidelines of the European Community's Council for Animal Experiments (86/609/EEC). Efforts were made to minimize the number of animals used and their suffering.

Quisqualic Acid and ABeta(25-35) Peptide Injections into the Nucleus Basalis and Drug Treatment

The quisqualic acid (Sigma Chemical Co., Milan, Italy; dissolved in phosphate buffer at the concentration of 0.12 M volume 0.5 μl) or 10 ug of ABeta(25-35) peptide (Bachem) dissolved PBS at the concentration of 10 μg/μl and aggregate at 37° for 2 h before injection volume 1 μl) was injected by means of an Hamilton microsyringe into the right NBM under chloral hydrate anaesthesia at the following stereotaxic coordinates: AP=−0.2, L=−2.8 from bregma and H=7 from the dura (Paxinos and Watson, 1982, Casamenti et al., 1998). Controlateral NBM were injected with PBS solution. The study was performed for 7 days after surgery. Rats were orally (ip) administered with a compound of the invention or vehicles with two administration, 24 h and 1 h before surgery and once daily for 7 days after lesioning. Last administration was performed 1 h before sacrifice.

Object Recognition Test

Object recognition was evaluated according to Ennanceur and Delacour (1988) and Scali et al., (1997). Briefly, the rats were placed in a grey polyvinylchloride arena (60×60×40 h cm) illuminated by a 50 W lamp suspended 50 cm above the arena. The objects to be discriminated were prisms, pyramids and cylinders made of plastic. The day before testing, rats were allowed to explore the arena for 2 min. On the day of the test, in the scopolamine protocol, a session of 2 trials separated by an intertrial interval of 240 min was carried out. In the first trial (acquisition trial, T1) two identical objects were presented in two opposite corners of the arena. The rats were left in the arena until criterion of 20 s of total exploration of the objects was reached. Exploration was defined as directing the nose at a distance <2 cm to the object and/or touching it with the nose. During the second trial (retention trial, T2) one of the objects presented in T1 was replaced by a new (differently shaped) object and the rats were left in the arena for 5 min. The times spent exploring the familiar (F) and the new object (N) were recorded separately and the difference between the two exploration times was taken. From one rat to the next, care was taken to avoid object and place preference by randomly changing the role of the objects (familiar or new object) and their position in the two opposite corners of the box during T2. Furthermore, in order to avoid olfactory stimuli the objects to be discriminated were cleaned carefully. In the time delay procedure, T2 was performed 24 h after T1 when a spontaneous decay of memory was presents in control rats. Drug's administrations were performed 30 min before the acquisition trial T1.

Immunohistochemistry

Under deep chloral hydrate anaesthesia, the rats were perfused transcardially with ice-cold paraformaldehyde solution (4% in phosphate-buffer, pH 7.4). The brains were postfixed for 4 h and cryoprotected in 18% sucrose solution for at least 48 h. Brains were cut in a cryostat throughout the injected area into 30 μm-thick coronal sections and placed in anti-freezer solution (phosphate-buffered saline containing 30% ethylene glycol and 30% glycerol) and stored at −20° C. until used for immunohistochemistry, according to the following schedule.

Day 1 ChAT (marker of cholinergic neurons, goat antiserum, Millipore, 1:200), was used as neurodegeneration marker and GFAP (marker of astrocytes, rabbit polyclonal antibody DAKO, 1:1000) and Iba-1 (marker of microglia, rabbit antibody, Wako, 1 1:500) or OX-42 (CD11b/c, marker of activated microglia, mouse antibody, BD Biosciences Pharmingen, 1:400) as neuroinflammation markers of astrocytes and microglia, respectively.

Secondary antibodies: biotinylated IgG (Vector Laboratories, Burlingame, Calif.), diluted 1:1000.

Immunohistochemical Procedure

Slices were processed as free-floating sections, briefly, the primary antibody was added at the appropriate dilution (in Blocking buffer: PBST with 0.5% BSA) and left overnight under mild agitation at room temperature. Then the corresponding biotinylated secondary antibody (in Blocking buffer: PBST with 0.1% BSA) was added to the slices and left 90 min at room temperature, under mild agitation, and then removed and washed with PBS. Bound antibody was visualized by using Vectastain ABC Kit (Vector Laboratories, Burlingame, Calif.) with DAB (Vector Laboratories, Burlingame, Calif.) as chromogen. The sections were mounted, counterstained with Ematossilin (Carlo Erba Reagents, Italy), dehydrated and coverslipped with mounting medium (Leica).

Immunohistochemical Marker Quantification

All immunohistochemical markers were quantified in the NMB area by the Aperio® digital pathology platform; briefly, 4-6 slides per animal were digitalized by using the scanner Scanscope CS (Aperio®), then the right and left NMB areas were manually identified for each slide creating a Region Of Interest (ROI) where specific macros of analysis were applied to quantify the signal. Each right striatum (injected with AAV9-Ex1-AcGFP-Q138) was compared with its contralateral (left) one (injected with AAV9-Ex1-AcGFP-Q17). Data from each slide were averaged on a per animal basis and the resulting values were used for statistical analysis.

ChAT was quantified as number of cells per area, as a single cell population. GFAP and Iba-1, were evaluated as positive pixel counts per area in the Region of Interest (ROI).

Formulation and Administration

Compounds under formula (A) are formulated preferably in admixture with a pharmaceutically acceptable carrier, excipient or the like. In general, it is preferable to administer the pharmaceutical composition in orally-administrable form, but certain formulations may be administered via a parenteral, intravenous, intramuscular, transdermal, buccal, subcutaneous, suppository, nasal or other route. One of ordinary skill in the art may modify the formulations within the teachings of the specification to provide numerous formulations for a particular route of administration without rendering the compositions of the present invention unstable or compromising their therapeutic activity.

In particular, the modification of the present compounds to render them more soluble in water or other vehicle, for example, may be easily accomplished by minor modifications (salt formulation, esterification, etc.) which are well within the ordinary skill in the art. It is also well within the routineer's skill to modify the route of administration and dosage regimen of a particular compound in order to manage the pharmacokinetics of the present compounds for maximum beneficial effect in patients.

In certain pharmaceutical dosage forms, the pro-drug form of the compounds, especially including ester and ether derivatives, as well as various salt forms of the present compounds, are preferred.

One of ordinary skill in the art will recognize how to readily modify the present compounds to pro-drug forms to facilitate delivery of active compounds to a targeted site within the host organism or patient.

The routineer also will take advantage of favourable pharmacokinetic parameters of the pro-drug forms, where applicable, in delivering the present compounds to a targeted site within the host organism or patient to maximize the intended effect of the compound.

Actual methods of preparing such dosage forms are known, or will be apparent, to those skilled in this art; for example, see Remington's Pharmaceutical Sciences, Mack Publishing Company, Easton, Pa., 15th Edition, 1975.

The composition or formulation to be administered will, in any event, contain a quantity of the active compound in an amount effective to alleviate the symptoms of the subject being treated.

While human dosage levels have yet to be optimized for the compounds of the invention, generally, a daily dose is from about 0.05 mg/kg to about 100 mg/kg of body weight.

The amount of active compound administered will, of course, be dependent on the subject and disease state being treated, the severity of the affliction, the manner and schedule of administration and the judgment of the prescribing physician.

For purposes of the present invention, a prophylactically or preventive effective amount of the compositions according to the present invention (i.e., an amount which substantially reduces the risk that a patient will either succumb to a disease state or condition or that the disease state or condition will worsen) falls within the same concentration range as set forth above for therapeutically effective amounts and is usually the same as a therapeutically effective amount.

In some embodiments of the present invention, one or more compounds of formula (A) are administered in combination with one or more other pharmaceutically active agents. The phrase “in combination”, as used herein, refers to agents that are simultaneously administered to a subject. It will be appreciated that two or more agents are considered to be administered “in combination” whenever a subject is simultaneously exposed to both (or more) of the agents.

Each of the two or more agents may be administered according to a different schedule; it is not required that individual doses of different agents be administered at the same time, or in the same composition. Rather, so long as both (or more) agents remain in the subject's body, they are considered to be administered “in combination”.

EXAMPLES

Experimental Section

All reagents and solvents were obtained commercially. Air and moisture sensitive liquid solutions were transferred via syringe. The course of reactions was followed by thin-layer chromatography (TLC) and/or liquid chromatography-mass spectrometry (HPLC-MS or UPLC-Ms). TLC analyses were performed on silica (Merck 60 F254) and spots revealed by UV visualisation at 254 nm and KMnO⁴ or ninhydrin stain.

Purifications by column chromatography were performed using silica cartridges Isolute flash Si or silica (Merck 60) or with flash chromatography purification instruments (Biotage). Compounds purities were above 90%.

All nuclear magnetic resonance spectra were recorded using a Bruker Avance AV 400 System (400.13 MHz for ¹H) equipped with BBI a probe.

ABBREVIATION

THF: Tetrahydrofuran

NH₄Cl: Ammonium chloride

AcOEt: Ethyl Acetate

Na₂SO₄: Sodium sulphate

HCl: Hydrochloric acid

DMF: N,N-dimethylformamide

NaH: Sodium Hydride

H₂O: Water

DCM: dichloromethane

NaOH: sodium hydroxide

K₂CO₃: potassium carbonate

NaHCO₃: sodium hydrogencarbonate

MeOH: methanol

EDC: 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride

DCE: 1, 2-dichloroethane

DIPEA: N,N-diisopropyl-N-ethylamine

NaCl: sodium chloride

K₃PO₄: Tripotassium phosphate

Pd₂ (dba)₃: Tris(dibenzylideneacetone)dipalladium (0)

Qphos: 1,2,3,4,5-Pentaphenyl-1′-(di-tert-butylphosphino)ferrocene

T₃P: Propylphosphonic Anhydride

EtOH: ethanol

Cs₂CO₃: Cesium carbonate

LiHMDS: lithium bis(trimethylsilyl)amide

H₂SO₄: Sulfuric acid

LDA: Lithium diisopropylamide

cHex: cyclohexane

NH₄OH: ammonium hydroxide

H₂: hydrogen

Pd/C: palladium on activated carbon

CDI: 1,1′-Carbonyldiimidazole

CH₃CN: Acetonitrile

Analytical Methods

Method c: Anaytical HPLC-MS were run using a Waters 2795 separation module equipped with a Waters Micromass ZQ (ES ionisation) and Waters PDA 2996, using a X-Bridge C18 3.5 μm 2.10×50 mm column. Gradient: 0.1% ammonia/water and acetonitrile with gradient 85/15 to 5/95 flow 0.8 ml/min over 5/10 minutes. Temperature: 40° C. UV Detection at 215 and 254 nm. ESI+ detection in the 80-1000 m/z range

Method d: Analytical UPLC-MS were run using a Acquity Waters UPLC with equipped with a Waters SQD (ES ionization) and Waters Acquity PDA detector, using a column BEH C18 1.7 μm, 2.1×5.00. Temperature: 40° C. UV Detection at 215 and 254 nm. ESI+ detection in the 80-1000 m/z range Gradient 0.1% ammonium bicarbonate/water and acetonitrile with a gradient 95/5 to 15/85 flow: 0.8 ml/min over 4 min.

Method e: Analytical UPLC-MS were run using a Acquity Waters UPLC with equipped with a Waters SQD (ES ionization) and Waters Acquity PDA detector, using a column BEH C18 1.7 μm, 2.1×5.00. Temperature: 40° C. UV Detection at 215 and 254 nm. ESI+ detection in the 80-1000 m/z range. Gradient 0.04% formic acid/95% water/5% acetonitrile and CH3CN with a gradient 95/5 to 0/100 flow: 0.8 ml/min over 4 minutes.

Method f: Analytical UPLC-MS were run using a Acquity Waters UPLC with equipped with a Waters SQD (ES ionization) and Waters Acquity PDA detector, using a column BEH C18 1.7 μm, 2.1×5.00. Temperature: 40° C. UV Detection at 215 and 254 nm. ESI+ detection in the 80-1000 m/z range. Gradient 0.1% formic acid/water and 0.1% formic acid/CH3CN with a gradient 95/5 to 5/95 flow: 0.6 ml/min over 3 minutes.

Preparative HPLC Method

Method a: Preparative HPLC was run using a Waters 2767 system with a binary gradient Module Waters 2525 pump and coupled to a Waters Micromass ZQ 25 (ES) or Waters 2487 DAD, using a X-Bridge C18 5 μm 19×150. Gradient 0.1% ammonia/water and methanol flow: 17 ml/min.

Method b: Preparative HPLC was run using a Waters 2767 system with a binary gradient Module Waters 2525 pump and coupled to a Waters MS3100 SQ or Waters 2487 DAD, using a X-Bridge C18 5 μm 19×150. Gradient 0.1% formic acid/water and 0.1% formic acid/methanol flow: 17 ml/min.

General Synthetic Procedures

General Procedure A1 for Carboxylation

To a solution of N,N-diisopropylamine (2.1 eq) in anhydrous THF (0.4 mL*mmol) cooled to −78° C., a solution of n-butyllithium (2.5 M in hexane, 2 eq) was added dropwise under an inert atmosphere. The mixture was stirred at −78° C. for one hour and then the desired methylpyridine (1 eq) was added. The reaction mixture was stirred at −78° C. for one hour and a solution of diethyl carbonate (1.2 eq) in THF (0.3 mL*mmol) was added. The reaction mixture was allowed to warm up to room temperature and left stirring overnight. The mixture was quenched with H₂O and extracted twice with AcOEt. The organic layer was collected, washed with saturated sodium chloride solution, dried over Na₂SO₄ and concentrated under reduced pressure. The crude product was purified by silica gel chromatography.

General Procedure A2 for Reformatsky-Negishi Coupling

To prepare the Reformatsky reagent Zinc dust (1.2 eq) was suspended in anhydrous THF under N₂ and Trimethylsilyl chloride 0.1 eq was added dropwise and the resulting suspension was refluxed for 1 h. Then bromoacetic acid tert-butyl ester (1.2 eq) was added dropwise and the resulting reaction mixture was refluxed for 2 h. The resulting Reformatsky reagent was added to a degassed suspension of Bromo-aryl or heteroaryl compound (1 eq), Q-phos (0.05 eq) and Palladium source (0.05 eq.) in anhydrous THF. The resulting reaction mixture was heated at 75° C. overnight. The reaction was worked up adding a saturated solution of NH₄Cl and AcOEt. The aqueous layer was extracted again with AcOEt and the resulting organic layers were combined, dried over Na₂SO₄ and concentrated in vacuo. The crude product was purified by silica gel chromatography.

General Procedure B1 for the Alkylation of Acid:

To a solution of heteroaryl-acetic acid (1 eq) in anhydrous THF cooled to −78° C., a solution of LiHMDS (1M 2.2 eq) in THF was added. The resulting mixture was stirred at −78° C. for 1 hour. Then 1-iodoproprane (1.1 eq) was added portionwise and the reaction mixture was allowed to warm up to room temperature and left stirring overnight. The reaction mixture was quenched with H₂O and extracted with AcOEt. The aqueous layer was separated; the solution was acidified to pH3 with 6N HCl and extracted with AcOEt three times. The organic phases were collected, dried over Na₂SO₄ and concentrated under reduced pressure. The crude product was purified by silica gel chromatography.

General Procedure B2 for Alkylation—Cyclization

Ethyl 2-(5-bromopyridin-3-yl)acetate (1 eq) was dissolved in DMF (5 mL*mmol); 18-crown-6 ether (0.05 eq) and NaH 60% dispersion in mineral oil (2.5 eq) were added and the mixture was stirred at room temperature for 30 minutes; dibromo alkane (1.1 eq) was added dropwise and reaction was stirred at room temperature for 5 h. NaOH 15% solution in H₂O (1.5 mL*mmol) was added and the mixture was stirred for 16 h at room temperature. H₂O was added and pH was adjusted to 3 with HCl 6N. Aqueous solution was extracted with DCM; organic phases were collected, dried over Na₂SO₄, filtered and evaporated. The crude product was purified by silica gel chromatography

General Procedure B3 for Alkylation

A solution of acid (1 eq) in dry THF (1.4 mL*mmol) was added dropwise to a solution of n-Butyllithium 1.6 M in n-hexane (2.2 eq) in THF (0.3 mL*mmol) at −78° C. The reaction was stirred at −78° C. in inert atmosphere for 2 h; then a solution of haloalkane (1.1 eq) in THF (0.6 mL*mmol) was added dropwise. Solution was allowed to warm up to room temperature and stirred for 16 h. H₂O was carefully added and mixture was diluted with AcOEt. Aqueous phase was collected, acidified to pH=1 with 6N HCl and extracted with AcOEt; organic layer was collected, dried over Na₂SO₄, filtered and evaporated under vacuum

General Procedure B4 for Fluoro Insertion

A solution of LiHMDS 1M in THF (1.1 eq) was diluted with THF (4.0 mL*mmol) and cooled to −78° C.; a solution of ethyl ester (1.0 eq) in the same solvent (2.0 mL*mmol) was added drop-wise. The mixture was stirred at 0° C. for 30 minutes and then cooled to −78° C. again. A solution in THF (4.0 mL*mmol) of N-fluorobenzene sulfonimide (1.3 eq) was added dropwise; the mixture was then warmed to room temperature and stirred for 12 hours. The reaction was quenched with NH₄Cl saturated aqueous solution, extracted with AcOEt and washed with H₂O. The organic layer was collected and the solvent was removed under reduce pressure. The crude product was purified by silica gel chromatography.

General Procedure B5 for Alkylation of Acid and Ester

Heteroaryl acetic acid ethyl ester (1 eq) was dissolved in DMF (2 mL*mmol), cesium carbonate (1.2 eq) and iodoalkane (1.1 eq) were added and the mixture was stirred at room temperature overnight. H₂O was added and crude extracted three times with AcOEt. Organic phases were combined, dried over Na₂SO₄ and concentrated in vacuo. The crude product was purified by silica gel chromatography.

General Procedure C1 for Phenol Alkylation

To a suspension of the desired phenol (1 eq) and K₂CO₃ (2 eq) in DMF, the desired alkyl bromide (4 eq) was added and the mixture was heated at 70° C. for 18 hours. _(H2O) was added and the mixture was extracted with AcOEt. The organic phase was collected and concentrated under reduced pressure. The crude product was purified by silica gel chromatography.

General Procedure D1 for Acid Hydrolysis

A solution of the desired ester in concentrated HCl (0.37 mmol/mL) was stirred at 100° C. for two hours. The mixture was concentrated under reduce pressure and the crude product was used in the next step without further purification.

General Procedure D2 for Acid Hydrolysis

To a solution of tert-butyl ester (1 eq) in DCM (10 mL*mmol), trifluoroacetic acid (1 mL*mmol) was added and the mixture was stirred at room temperature for three days. The mixture was concentrated under reduced pressure, then was diluted with DCM and extracted with NaHCO₃ saturated solution. The aqueous layer was separated, acidified to pH3 with HCl 1N and extracted with DCM. The organic phase was separated, dried over Na₂SO₄ and concentrated under reduced pressure, affording the title compound.

General Procedure D3 for Basic Hydrolysis

To a solution of ester (1 eq) in MeOH (7.5 mL*mmol), a solution of 2N NaOH (7.5 mL*mmol) was added and the mixture was stirred at room temperature for 3 hours. The solvent was removed under reduced pressure, the residue was suspended in H₂O and the mixture was acidified with 1N HCl to pH3. The aqueous phase was extracted with DCM and the organic layer was collected and dried over Na₂SO₄. The title compound was obtained without further purification.

General Procedure E1 for the Amide Coupling with Thionyl Chloride:

To a solution of carboxylic acid (1 eq) in 1,2-dichloroethane (4.3 mL*mmol) thionyl chloride (1.2 eq) and catalytic amount of DMF were added and the mixture was stirred at 60° C. for 4 hours. Then the mixture was allowed to cool down to room temperature and the desired amine (1.2 eq) and DIPEA (3 eq) were added. The mixture was stirred at room temperature overnight then washed with saturated NaHCO₃ solution, the organic layer collected and the solvent was removed under reduce pressure. The crude product was purified by silica gel chromatography.

General Procedure E2 for Amide Coupling with EDC and 1-Hydroxybenzotriazole Hydrate

To a solution of acid (1 eq) in DMF (3 mL*mmol), amine (1.1 eq), 1-hydroxybenzotriazole hydrate (0.36 eq) and EDC (1.5 eq) were added. The mixture was stirred at room temperature for one hour. NaHCO₃ saturated solution was added and the mixture was extracted with DCM. The combined organic extracts were washed with saturated NaCl solution, dried over Na₂SO₄ and evaporated. The crude product was purified by silica gel chromatography.

General Procedure E3 for the Amide Coupling with N-Bromosuccinimide Triphenylphosphine:

To a solution of triphenylphosphine (1.6 eq) in DCM (1 ml*mmol of carboxylic acid) cooled at 0° C., N-bromosuccinimide (1.6 eq) was added and the mixture left at 0° C. for 30 minutes. The desire carboxylic acid (1 eq) was added and the reaction was allowed to warm up to room temperature and lest stirring for 45 minutes. Amine (2.5 eq) was added and the mixture was left stirring for 18 hours at room temperature. The mixture was washed with 1N HCl solution and NaHCO₃ saturated solution. The organic phase was collected and the solvent was removed under reduced pressure. The crude product was purified by silica gel chromatography.

General Procedure E4 for the Amide Coupling with T3P

To a solution of carboxylic acid (1 eq) and amine (1 eq) in AcOEt, DIPEA (2 eq) was added and solution cooled to 0° C. T3P 50% solution in AcOEt (1.5 eq) was added and reaction was stirred for 12 h at room temperature. NaHCO₃ saturated solution was added, organic layer was separated, dried over Na₂SO₄, filtered and evaporated. The crude product was purified by silica gel chromatography.

General Procedure F1 for Amide Coupling of Ester

To a solution of acid (1 eq 0.12 g, 0.47 mmol) in DMF (3 mL*mmol), amine (1.1 eq), 1-hydroxybenzotriazole hydrate (0.36 eq) and EDC (1.5 eq) were added. The mixture was stirred at room temperature for one hour. NaHCO₃ saturated solution was added and the mixture was extracted with DCM. The combined organic extracts were washed with saturated NaCl solution, dried over Na₂SO₄ and evaporated. The crude product was purified by silica gel chromatography.

General Procedure I for Nitrile Alcoholysis

To a solution of EtOH (2 mL*mmol of nitrile) H₂SO₄ concentrated (0.76 mL*mmol of nitrile) was added dropwise and the desired nitrile (1 eq) was added portion wise. The solution was stirred at 100° C. for three hours. The mixture was added dropwise to a solution of NaHCO₃ (3.00 g*mmol of nitrile) in H₂O (7.5 mL*mmol of nitrile) and it was extracted twice with DCM. The organic layer were collected, dried and evaporated, affording the desired compound.

General Procedure C1 for Phenol Alkylation

To a suspension of the desired phenol (1 eq) and K₂CO₃ (2 eq) in DMF, the desired alkyl bromide (4 eq) was added and the mixture was heated at 70° C. for 18 hours. H₂O was added and the mixture was extracted with AcOEt. The organic phase was collected and concentrated under reduced pressure. The crude product was purified by silica gel chromatography.

General Procedure J1 for Alkylation

To a solution of N-heterocycle (1 eq) in DMF (2 ml*mmol), NaH (60% in mineral oil, 1.2 eq) was added and the mixture was stirred at room temperature for 30 minutes. 2-Bromo-alkanoic acid ethyl ester (1.1 eq) was added and the reaction was left stirring at room temperature overnight. Saturated NaCl solution was added and the mixture was extracted with DCM. The organic phase was collected, dried over Na₂SO₄ and concentrated under reduced pressure. The crude product was purified by silica gel chromatography.

General Procedure J2 for Alkylation

A suspension of N-heterocycle (1 eq) and K₂CO₃ (2 eq) in acetone (4 mL*mmol) was heated at 55° C. for 10 minutes and then was allowed to cool down to room temperature. 2-Bromo-alkanoic acid ethyl ester (1.1 eq) was the added and the mixture was heated at 55° C. for 18 hours. The solvent was removed under reduced pressure and the crude product was suspended in DCM and washed with H₂O. The organic phase was collected, dried over Na₂SO₄ and concentrated under reduced pressure.

General Procedure O for Suzuki Coupling

Ester/Acid (1 eq) was dissolved in degassed dioxane (4 mL*mmol), boronic acid or ester (1 eq), K₃PO₄ (1.7 eq), phosphine (0.02 eq), Pd₂(dba)₃ (0.01 eq) were added then degassed H₂O (0.5 mL*mmol) was added and reaction mixture was heated at 100° C. in a pressure tube for 16 h. AcOEt and NaCl saturated solution were added. Organic phase was collected and evaporated. The crude product was purified by silica gel chromatography.

Example 1: 2-(5-Bromo-pyridin-3-yl)-pentanoic Acid (5-bromo-pyridin-2-yl)-amide

2-(5-Bromo-pyridin-3-yl)-pentanoic Acid

To a solution of (5-Bromo-pyridin-3-yl)-acetic acid (2.00 g, 9.2 mmol) in anhydrous THF (20 mL) cooled to −78° C., a solution of LiHMDS (1M, 20 mmol) in THF was added. The resulting mixture was stirred at −78° C. for 1 hour. Then 1-iodo-proprane (1.70 g, 10.2 mmol) was added portionwise and the reaction mixture was allowed to warm up to room temperature and left stirring overnight. The reaction mixture was quenched with H₂O and extracted with AcOEt. The aqueous layer was separated; the solution was acidified to pH3 with HCl 6N and extracted with AcOEt three times. The organic phases were collected, dried over Na₂SO₄ and concentrated under reduced pressure. The crude product was purified by silica gel chromatography (cHex/AcOEt 1/1) to afford the title compound (1.2 g, 50%).

C₁₀H₁₂BrNO₂ Mass (calculated) [258.12]; (found) [M+H]⁺=269.

2-(5-Bromo-pyridin-3-yl)-pentanoic Acid (5-bromo-pyridin-2-yl)-amide

To a solution of 2-(5-bromo-pyridin-3-yl)-pentanoic acid (0.12 g, 0.47 mmol) in DCE (2 mL) thionyl chloride (0.08 g, 0.56 mmol) and catalytic amount of DMF were added and the mixture was stirred at 60° C. for four hours. Then the mixture was allowed to cool down to room temperature and 5-bromo-pyridin-2-ylamine (0.10 g, 0.59 mmol) and DIPEA (0.18 g, 1.395 mmol) were added. The mixture was stirred at room temperature overnight then washed with sodium bicarbonate saturated solution, the organic layer collected and the solvent was removed under reduce pressure. The crude product was purified by silica gel chromatography (cHex/AcOEt 1/1), to afford the title compound (0.05 g, 25%).

¹H NMR (400 MHz, Chloroform-d3) δ 8.62 (d, J=2.1 Hz, 1H), 8.49 (d, J=2.1 Hz, 1H), 8.31 (d, J=2.4 Hz, 1H), 8.14 (d, J=8.8 Hz, 1H), 7.98-7.91 (m, 2H), 7.81 (dd, J=8.8, 2.4 Hz, 1H), 3.47 (t, J=7.5 Hz, 1H), 2.23-2.12 (m, 1H), 1.87-1.75 (m, 1H), 1.44-1.24 (m, 2H), 0.96 (t, J=7.3 Hz, 3H).

C₁₅H₁₅Br₂N₃O, Calculated [413.11], found [M+H⁺] 414, RT=1.74 (method f).

Example 2: 2-(5-Bromo-pyridin-3-yl)-hexanoic Acid (5-bromo-3-fluoro-pyridin-2-yl)-amide

2-(5-Bromo-pyridin-3-yl)-hexanoic Acid

The title compound was obtained following general procedure for alkylation B1 and starting from (5-bromo-pyridin-3-yl)-acetic acid and 1-Iodo-butane (1.82 g, 51%).

C₁₁H₁₄BrNO₂ Mass (calculated) [272]; (found) [M+H]⁺=274.

5-Bromo-3-fluoro-pyridin-2-ylamine

To a solution of 3-fluoro-pyridin-2-ylamine (0.30 g, 2.68 mmol) in acetonitrile (15 mL), in inert atmosphere, N-bromosuccinimide (0.48 g, 2.68 mmol) was added. The mixture was stirred for 4 hours. Solvent was removed under reduced pressure and the crude product was purified by silica gel chromatography (cHex/AcOEt 66/34) to give the title compound (0.46 g, 89%).

C₅H₄BrFN₂ Mass (calculated) [191]; (found) [M+H]⁺=193.

2-(5-Bromo-pyridin-3-yl)-hexanoic Acid (5-bromo-3-fluoro-pyridin-2-yl)-amide

The title compound was obtained following general procedure E1 for amide coupling and starting from 2-(5-bromo-pyridin-3-yl)-hexanoic acid and 5-bromo-3-fluoro-pyridin-2-ylamine, (0.10 g, 34%).

¹H NMR (400 MHz, Chloroform-d3) δ 8.60 (d, J=2.2 Hz, 1H), 8.49 (d, J=2.2 Hz, 1H), 8.27 (d, J=2.0 Hz, 1H), 7.97 (t, J=2.2 Hz, 1H), 7.78 (s, 1H), 7.64 (dd, J=9.0, 2.0 Hz, 1H), 3.90 (bs, 1H), 2.28-2.14 (m, 1H), 1.88-1.74 (m, 1H), 1.46-1.18 (m, 4H), 0.89 (t, J=7.1 Hz, 3H). C₁₆H₁₆Br₂FN₃O, Calculated [445.12], found [M+H⁺], 2Br pattern 446, RT=1.64 (method f).

Example 3: N-(5-Bromo-6-fluoro-pyridin-2-yl)-2-(5-bromo-pyridin-3-yl)-3-methyl-butyramide

2-(5-Bromo-pyridin-3-yl)-3-methyl-butyric Acid

The title compound was prepared following general procedure B1 for the alkylation of acid. (1.80 g, 61%).

Mass (calculated C₁₀H₁₂BrNO₂ [258]; found [M+1]=258-260 bromine pattern.

N-(5-Bromo-6-fluoro-pyridin-2-yl)-2-(5-bromo-pyridin-3-yl)-3-methyl-butyramide

Amide coupling was performed with thionyl chloride following the procedure E4. The crude product was purified by silica gel chromatography eluting (CHex/AcOEt 0-40%) to give title compound (0.09 g, 36%).

¹H NMR (400 MHz, Chloroform-d3) δ 8.62 (d, J=2.2 Hz, 1H), 8.46 (d, J=1.9 Hz, 1H), 8.05-7.97 (m, 2H), 7.93 (t, J=8.5 Hz, 1H), 7.84 (bs, 1H), 2.98 (d, J=10.2 Hz, 1H), 2.52-2.38 (m, 1H), 1.12 (d, J=6.5 Hz, 3H), 0.79 (d, J=6.6 Hz, 3H). C15H14N3OFBr2, Calculated [431.10], found [M+H⁺], 432, RT=1.79 (method f).

Example 4: 1-(5-Bromo-pyridin-3-yl)-cyclobutanecarboxylic Acid (5-bromo-pyridin-2-yl)-amide

1-(5-Bromo-pyridin-3-yl)-cyclobutane carboxylic Acid

Ethyl 2-(5-bromopyridin-3-yl)acetate (1.0 g, 4.09 mmol, 1 eq) was dissolved in DMF (20 mL); 18-crown-6 ether (0.054 g, 0.205 mmol, 0.05 eq) and NaH 60% dispersion in mineral oil (0.41 g, 10.2 mmol, 2.5 eq) were added and the mixture was stirred at room temperature for 30 minutes; 1,3-dibromopropane (0.46 mL, 4.50 mmol, 1.1 eq) was added dropwise and reaction was stirred at room temperature for 5 h. NaOH 15% solution in H₂O (10 mL) were added and the mixture was stirred for 16 h at room temperature. H₂O was added and pH was adjusted to pH=3 with HCl 6N. Aqueous solution was extracted with DCM (2×20 mL), organic phases were collected, dried over Na₂SO₄, filtered and evaporated. The crude product was purified by silica gel chromatography (cHex/AcOEt 5%-60%) to give the title compound (0.38 g, 37% over two steps).

C₁₀H₁₀BrNO₂Mass (calculated) [256]; found [M+1]=256-258 bromine pattern.

1-(5-Bromo-pyridin-3-yl)-cyclobutanecarboxylic Acid (5-bromo-pyridin-2-yl)-amide

Amide coupling on 1-(5-bromo-pyridin-3-yl)-cyclobutane carboxylic acid and 5-bromo-pyridin-2-ylamine was performed using general procedure E2 to give the title compound (0.015 g, 11%).

¹H NMR (400 MHz, Chloroform-d3) δ 8.66-8.56 (m, 2H), 8.29-8.24 (m, 1H), 8.19-8.12 (m, 1H), 7.88-7.76 (m, 2H), 7.58 (s, 1H), 3.01-2.89 (m, 2H), 2.63-2.51 (m, 2H), 2.24-1.93 (m, 2H).

C₁₅H₁₃N₃OBr₂, Calculated [411.09], found [M+H⁺], 2Br pattern 412, RT=1.61 (method f).

Example 5: 1-(5-Bromo-pyridin-3-yl)-cyclopentanecarboxylic Acid (5-bromo-pyrazin-2-yl)-amide

1-(5-Bromo-pyridin-3-yl)-cyclopentanecarboxylic Acid

Starting from 1-(5-bromo-pyridin-3-yl)-acetic acid ethyl ester, the title compound was synthesised using the general procedure B2 for cyclization, followed by basic hydrolysis (D3) (0.66 g, 59% over two steps).

Mass (calculated) C₁₁H₁₂BrNO₂ [270]; found [M−1]=270-272 bromine pattern.

1-(5-Bromo-pyridin-3-yl)-cyclopentanecarboxylic Acid (5-bromo-pyrazin-2-yl)-amide

Amide coupling was performed with thionyl chloride following the procedure E1 to give the title compound (0.021 g, 9%).

¹H NMR (400 MHz, Chloroform-d) δ 9.31 (d, J=1.7 Hz, 1H), 8.62 (dd, J=10.7, 2.1 Hz, 2H), 8.28 (d, J=1.7 Hz, 1H), 7.86 (t, J=2.1 Hz, 1H), 7.53 (s, 1H), 2.71-2.55 (m, 2H), 2.17-2.03 (m, 2H), 1.99-1.73 (m, 4H).

C₁₅H₁₄N₄OBr₂, Calculated [426.11], found [M+H⁺], 2Br pattern, 427, RT=1.61 (method f).

Example 6: 1-(5-Chloro-pyridin-3-yl)-cyclobutanecarboxylic Acid (5-chloro-pyridin-2-yl)-amide

(5-Chloro-pyridin-3-yl)-acetic Acid tert-butyl ester

The title compound was synthesized from 3-bromo-5-chloropyridine using general procedure A2 for alkylation (8.20 g, 77%).

C₁₁H₁₄ClNO₂Mass (calculated) [227]; found [M+1]=228-230 chlorine pattern.

1-(5-Chloro-pyridin-3-yl)-cyclobutanecarboxylic Acid tert-butyl ester

The title compound was prepared using the general procedure B2 for cyclization starting from (5-Chloro-pyridin-3-yl)-acetic acid tert-butyl ester and 1,3-diiodopropane (0.47 g, 37%).

C₁₄H₁₈ClNO₂Mass (calculated) [267]; found [M+1]=268-270 chlorine pattern.

1-(5-Chloro-pyridin-3-yl)-cyclobutanecarboxylic Acid

The acid was obtained from 1-(5-Chloro-pyridin-3-yl)-cyclobutanecarboxylic acid tert-butyl ester using general procedure D2 for acid hydrolysis (0.33 g, quant.).

C₁₀H₁₀ClNO₂Mass (calculated) [211]; found [M+1]=268-270 chlorine pattern.

1-(5-Chloro-pyridin-3-yl)-cyclobutanecarboxylic Acid (5-chloro-pyridin-2-yl)-amide

Starting from 1-(5-Chloro-pyridin-3-yl)-cyclobutanecarboxylic acid and 5-bromo-pyridin-2-ylamine amide coupling was performed with thionyl chloride following the procedure E1 To give the title compound (0.05 g, 4%).

¹H NMR (400 MHz, Chloroform-d3) δ 8.56 (d, J=2.2 Hz, 1H), 8.53 (d, J=2.2 Hz, 1H), 8.20 (d, J=8.9 Hz, 1H), 8.17 (d, J=2.4 Hz, 1H), 7.70 (t, J=2.2 Hz, 1H), 7.67 (dd, J=8.9, 2.4 Hz, 1H), 7.58 (s, 1H), 3.03-2.90 (m, 2H), 2.64-2.51 (m, 2H), 2.25-2.09 (m, 1H), 2.09-1.94 (m, 1H).

C₁₅H₁₃N₃OCl₂, Calculated [322.19], found [M+H⁺], 322, RT=1.53 (method f).

Example 7: 1-(6-Chloro-5-cyano-pyridin-3-yl)-cyclobutanecarboxylic Acid (5-chloro-pyridin-2-yl)-amide

(6-Chloro-5-cyano-pyridin-3-yl)-acetic Acid tert-butyl ester

The title compound was synthesized from 5-Bromo-2-chloronicotinonitrile using general procedure A2 for alkylation. (1.60 g, 35%).

Mass (calculated) C₁₂H₁₃ClN₂O₂ [252]; found [M+1]=253.

1-(6-Chloro-5-cyano-pyridin-3-yl)-cyclobutanecarboxylic Acid

The title compound was prepared using the general procedure B2 for cyclization, followed by acid hydrolysis using general procedure D2. (0.18 g, 30%).

Mass (calculated) C₁₁H₉ClN₂O₂ [236]; found [M+1]=237.

1-(6-Chloro-5-cyano-pyridin-3-yl)-cyclobutanecarboxylic Acid (5-chloro-pyridin-2-yl)-amide

Amide coupling was performed following the procedure E1, starting from 1-(6-chloro-5-cyano-pyridin-3-yl)-cyclobutanecarboxylic acid and acid (5-chloro-pyridin-2-yl)-amine to give the title compound (0.066 g, 50%).

¹H NMR (400 MHz, Chloroform-d3) δ 8.64 (d, J=2.5 Hz, 2H), 8.24-8.07 (m, 2H), 8.03 (d, J=2.5 Hz, 1H), 7.76 (s, 1H), 7.69 (dd, J=9.0, 2.5 Hz, 1H), 3.17-2.71 (m, 2H), 2.64-2.29 (m, 2H), 2.36-1.92 (m, 2H).

C₁₆H₁₂N₄OCl₂, Calculated [347.20], found [M+H⁺], 347, RT=1.59 (method f).

Example 8: 2-(6-Chloro-5-cyano-pyridin-3-yl)-pentanoic Acid (5-chloro-pyrazin-2-yl)-amide

(6-Chloro-5-cyano-pyridin-3-yl)-acetic Acid tert-butyl ester

The title compound was synthesized following the general procedure A2 starting from 5-bromo-2-chloro-nicotinonitrile. The crude product was purified by silica gel chromatography (cHex/AcOEt gradient) to give the title compound (0.85 g, 75% y).

¹H NMR (400 MHz, CDCl3) δ 8.47 (s, 1H), 7.97 (s, 1H), 3.58 (s, 2H), 1.46 (s, 9H).

2-(6-Chloro-5-cyano-pyridin-3-yl)-pentanoic Acid tert-butyl ester

Alkylation was performed following the general procedure B1 starting from (6-Chloro-5-cyano-pyridin-3-yl)-acetic acid tert-butyl ester, to give the title compound (0.60 g, 60% y).

C₁₅H₁₉ClN₂O₂Mass (calculated) [294]; (found) [M+H]⁺=295.

2-(6-Chloro-5-cyano-pyridin-3-yl)-pentanoic Acid

The title compound was synthesized following the general procedure D2 starting from 2-(6-Chloro-5-cyano-pyridin-3-yl)-pentanoic acid tert-butyl ester; (0.12 g, 98% y). C₁₁H₁₁ClN₂O₂ Mass (calculated) [238]; (found) [M+H]⁺=239.

2-(6-Chloro-5-cyano-pyridin-3-yl)-pentanoic Acid (5-chloro-pyrazin-2-yl)-amide

The title product was synthesized following the general procedure E2 starting from 2-(6-Chloro-5-cyano-pyridin-3-yl)-pentanoic acid and 5-Chloro-pyrazin-2-ylamine, (0.10 g, 53%).

¹H NMR (400 MHz, Chloroform-d3) δ 9.30 (s, 1H), 8.56 (d, J=2.3 Hz, 1H), 8.27 (s, 1H), 8.18 (d, J=2.3 Hz, 1H), 7.92 (s, 1H), 3.59 (t, J=7.6 Hz, 1H), 2.27-2.12 (m, 1H), 1.90-1.80 (m, 1H), 1.46-1.19 (m, 2H), 0.98 (t, J=7.3 Hz, 3H).

C₁₅H₁₃N₅OCl₂, Calculated [350.20], found [M+H⁺], 2 Cl pattern 350-352, RT=1.60 (method f).

Example 9: 2-(6-Chloro-5-fluoro-pyridin-3-yl)-pentanoic Acid (5-bromo-pyrazin-2-yl)-amide

(6-Chloro-5-fluoro-pyridin-3-yl)-acetic Acid tert-butyl ester

The title compound was synthesized following the general procedure A2 starting from 5-bromo-2-chloro-3-fluoro-pyridine. The crude product was purified by silica gel chromatography (cHex/AcOEt gradient) to give (6-Chloro-5-fluoro-pyridin-3-yl)-acetic acid tert-butyl ester (0.36 g, 30%).

C₁₁H₁₃ClFNO₂Mass (calculated) [245]; (found) [M+H]⁺=246.

2-(6-Chloro-5-fluoro-pyridin-3-yl)-pentanoic Acid tert-butyl ester

The title compound was synthesized following the general procedure B1 starting from (6-Chloro-5-fluoro-pyridin-3-yl)-acetic acid tert-butyl ester. The crude product was purified by silica gel chromatography (cHex/AcOEt gradient) to give 2-(6-Chloro-5-fluoro-pyridin-3-yl)-pentanoic acid tert-butyl ester (0.17 g, 56%).

C₁₄H₁₉ClFNO₂Mass (calculated) [287]; (found) [M+H]⁺=288.

2-(6-Chloro-5-fluoro-pyridin-3-yl)-pentanoic Acid

The title compound was synthesized following the general procedure D2 starting from (2-(6-Chloro-5-fluoro-pyridin-3-yl)-pentanoic acid tert-butyl ester (0.16 g, quant.). C₁₀H₁₁ClFNO₂Mass (calculated) [231]; (found) [M+H]⁺=232.

2-(6-Chloro-5-fluoro-pyridin-3-yl)-pentanoic Acid (5-bromo-pyrazin-2-yl)-amide

The title compound was synthesized following the general procedure E1 starting from 2-(6-Chloro-5-fluoro-pyridin-3-yl)-pentanoic acid and 5-Bromo-pyrazin-2-ylamine (0.03 g, 33%).

¹H NMR (400 MHz, Methanol-d4) δ 9.20 (d, J=1.5 Hz, 1H), 8.45 (d, J=1.5 Hz, 1H), 8.23 (d, J=1.9 Hz, 1H), 7.84 (dd, J=9.4, 1.9 Hz, 1H), 3.90 (dd, J=8.3, 7.0, Hz, 1H), 2.20-2.06 (m, 1H), 1.86-1.72 (m, 1H), 1.47-1.19 (m, 2H), 0.96 (t, J=7.4 Hz, 3H).

C₁₄H₁₃N₄OFClBr, Calculated [387.63], found [M+H⁺], Cl—Br pattern 389, RT=1.77 (method f).

Example 10: 2-(5-Bromo-pyridin-3-yl)2-Fluoro-pentanoic Acid (5-bromo-pyridin-2-yl) amide

2-(5-Bromo-pyridin-3-yl)-pentanoic Acid

To a solution of (5-Bromo-pyridil-3-yl) acetic acid (2.0 g, 9.3 mmol, 1 eq) in anhydrous THF cooled to −78° C., a solution of LiHMDS (20.4 mL, 20.4 mmol, 2.2 eq) in THF was added. The resulting mixture was stirred at −78° C. for 1 hour. Then 1-iodopropane (1.0 mL, 10.2 mmol, 1.1 eq) was added portionwise and the reaction mixture was allowed to warm up to room temperature and left stirring overnight. The reaction mixture was quenched with H₂O and extracted with AcOEt. The aqueous layer was separated; the solution was acidified to pH=3 with 6N HCl and extracted with AcOEt. The organic phases were collected, dried over Na₂SO₄ and concentrated under reduced pressure. The crude product was purified by silica gel chromatography (cHex: AcOEt 92:8 to 34:66) to give the title compound (1.2 g, 50%).

C₁₀H₁₂BrNO₂ mass (calculated) [258]; (found) [M+H]⁺=259 m/z.

2-(5-Bromo-pyridin-3-yl)-pentanoic ethyl ester

To a solution of 2-(5-Bromo-pyridin-3-yl)-pentanoic acid (1.50 g, 5.8 mmol, 1 eq) in EtOH (10 mL), H₂SO₄ (0.5 mL, 2.6 eq, 15.2 mmol) was added and the mixture was stirred at 85° C. for 12 hours. Then the mixture was allowed to cool to room temperature. The mixture was concentrated under reduced pressure, dissolved in DCM and washed with sodium bicarbonate saturated solution. The organic layer was collected and the solvent was removed under reduce pressure to give the desired product employed in the next step without further purification (1.5 g, 88%).

C₁₂H₁₆BrNO₂ mass (calculated) [286]; (found) [M+H]⁺=287 m/z.

2-(5-Bromo-pyridin-3-yl)-2-fluoro-pentanoic ethyl ester

A solution of LiHDMS (1M in THF, 0.58 mL, 1.1 eq) was diluted with THF (2.0 mL) and cooled to −78° C.; a solution of the 2-(5-Bromo-pyridin-3-yl)-pentanoic ethyl ester (0.15 g, 0.52 mmol, 1.0 eq) in the same solvent (1.0 mL) was added dropwise. The mixture was stirred at 0° C. for 30 minutes and then cooled to −78° C. again. A solution in THF (2.0 mL) of N-fluorobenzene sulfonimide (0.22 g, 0.68 mmol, 1.3 eq) was added dropwise; the mixture was then warmed to room temperature and stirred for 12 hours. The reaction was quenched with NH₄Cl saturated aqueous solution, extracted with AcOEt and washed with H₂O. The organic layer was collected and the solvent was removed under reduce pressure. The crude product was purified by silica gel chromatography (cHex: AcOEt 100:0 to 80:20) give the desired product as an orange oil (0.10 g, 63%). C₁₂H₁₅BrFNO₂ mass (calculated) [304]; (found) [M+H]⁺=305 m/z.

2-(5-Bromo-pyridin-3-yl)-2-fluoro-pentanoic carboxylic Acid

To a solution of 2-(5-Bromo-pyridin-3-yl)-2-fluoro-pentanoic ethyl ester (0.48 g, 1.6 mmol, 1 eq) in MeOH (3 mL), a solution of 2N NaOH (3 mL, 6 mmol, 4 eq) was added and the mixture was stirred at room temperature for 3 hours. The solvent was removed under reduced pressure, the residue was suspended in H₂O and the mixture was acidified with 1N HCl to pH=3. The aqueous phase was extracted with DCM and the organic layer was collected and dried over Na₂SO₄. The title compound was isolated without further purification (0.41 g, 95%).

C₁₀H₁₁BrFNO₂ Mass (calculated) [276]; (found) [M+H]⁺=277 m/z.

2-(5-Bromo-pyridin-3-yl)2-Fluoro-pentanoic Acid (5-bromo-pyridin-2-yl)-amide

To a solution of 2-(5-Bromo-pyridin-3-yl)-2-fluoro-pentanoic carboxylic acid (0.12 g, 0.44 mmol, 1 eq) in DMF (1.5 mL), 5-Bromo-2-aminopyridine (0.08 g, 0.48 mmol, 1.1 eq), 1-hydroxybenzotriazole hydrate (0.02 g, 0.13 mmol, 0.3 eq) and EDC (0.10 g, 0.52 mmol, 1.2 eq) were added. The mixture was stirred at room temperature for one hour. NaHCO₃ saturated solution was added and the mixture was extracted with DCM. The combined organic extracts were washed with saturated NaCl solution, dried over Na₂SO₄ and evaporated. The crude product was purified by silica gel chromatography (cHex: AcOEt 100:0 to 77:23) to give the title compound (0.07 g, 38%).

¹H NMR (400 MHz, Chloroform-d3) δ 8.76-8.68 (m, 2H), 8.61 (d, J=2.3 Hz, 1H), 8.29 (d, J=2.4 Hz, 1H), 8.05 (d, J=8.8 Hz, 1H), 8.01 (t, J=2.3 Hz, 1H), 7.76 (dd, J=8.8, 2.4 Hz, 1H), 2.42-2.21 (m, 1H), 2.18-1.99 (m, 1H), 1.47-1.31 (m, 2H), 0.89 (t, J=7.4 Hz, 3H).

C₁₅H₁₄Br₂FN₃O, Calculated [431.10], found [M+H⁺], 432, RT=2.35 (method e).

Example 11: 2-(5-Bromo-pyridin-3-yl)-2-methyl-pentanoic Acid (5-bromo-pyrazin-2-yl)-amide

2-(5-Bromo-pyridin-3-yl)-propionic Acid

(5-Bromo-pyridin-3-yl)-acetic acid was alkylated with iodomethane using general procedure B1 for the alkylation of acid to give the title product (0.6 g, 61%).

C₈H₈BrNO₂Mass (calculated) [230]; found [M+1]=230-232 bromine pattern.

2-(5-Bromo-pyridin-3-yl)-2-methyl-pentanoic Acid

2-(5-Bromo-pyridin-3-yl)-propionic acid was alkylated using general procedure B1 for the alkylation, heating at 50° C. to give the title compound (0.1 g, 28%). C₁₁H₁₄BrNO₂Mass (calculated) [272]; found [M+1]=272-274 bromine pattern.

2-(5-Bromo-pyridin-3-yl)-2-methyl-pentanoic Acid (5-bromo-pyrazin-2-yl)-amide

Amide coupling was performed with thionyl chloride following the procedure E1, starting from 2-(5-bromo-pyridin-3-yl)-2-methyl-pentanoic acid and 5-bromo-pyrazin-2-yl)-amine to give the title compound after preparative HPLC in basic condition (0.02 g, 10%).

¹H NMR (400 MHz, DMSO-d6) δ 10.45 (s, 1H), 9.10 (d, J=2.1 Hz, 1H), 8.61-8.53 (m, 2H), 8.44 (d, J=2.1 Hz, 1H), 7.91 (t, J=2.1 Hz, 1H), 2.16-2.03 (m, 1H), 2.01-1.83 (m, 1H), 1.57 (s, 3H), 1.20-1.02 (m, 2H), 0.85 (t, J=7.1 Hz, 3H).

C₁₅H₁₆N₄OBr₂, Calculated [428.1], found [M+H⁺], 2Br pattern 429, RT=1.66 (method f).

Example 12: 2-(6-Bromo-pyridin-2-yl)-pentanoic Acid (5-bromo-pyrazin-2-yl)-amide

(6-Bromo-pyridin-2-yl)-acetic Acid ethyl ester

To a solution of N,N-diisopropylamine (1.85 g, 18.31 mmol) in anhydrous THF (7 mL) cooled to −78° C., a solution of n-butyllithium (2.5 M in hexane, 17.44 mmol) was added drop wise under inert atmosphere. The mixture was stirred at −78° C. for one hour and then 2-bromo-6-methylpyridine (1.5 g, 8.7 mmol). The reaction mixture was stirred at −78° C. for one hour and a solution of diethyl carbonate (1.23 g, 10.46 mmol) in THF (3 mL) was added. The reaction mixture was allowed to warm up to room temperature and left stirring overnight. The mixture was quenched with H₂O and extracted twice with AcOEt. The organic layer was collected, washed with saturated sodium chloride solution, dried over Na₂SO₄ and concentrated under reduced pressure. The crude was purified by chromatography on silica gel (cHex/AcOEt 70/30) to give the title compound as an yellow oil (1.16 g, 55%).

C₉H₁₀BrNO₂ Mass (calculated) [244]; (found) [M+H]⁺=246.

2-(6-Bromo-pyridin-2-yl)-pentanoic Acid ethyl ester

The title compound was obtained following the general procedure B1 and starting from (6-bromo-pyridin-2-yl)-acetic acid ethyl ester (0.85 g, 63%).

C₁₂H₁₆BrNO₂ Mass (calculated) [286]; (found) [M+H]⁺=288.

2-(6-Bromo-pyridin-2-yl)-pentanoic Acid

To a solution of 2-(6-bromo-pyridin-2-yl)-pentanoic acid ethyl ester (0.40 g, 1.4 mmol) in MeOH (3 mL), a solution of 2N NaOH (3 ml) was added and the mixture was stirred at room temperature for 3 hours. The solvent was removed under reduced pressure, the crude product was suspended in H₂O and the mixture was acidified with 1N HCl to pH3. The aqueous phase was extracted with DCM and the organic layer was collected and dried over Na₂SO₄. The title compound was obtained in quantitative yield without further purification.

C₁₀H₁₂BrNO₂ Mass (calculated) [258]; (found) [M+H]⁺=260.

2-(6-Bromo-pyridin-2-yl)-pentanoic Acid (5-bromo-pyrazin-2-yl)-amide

To a solution of 2-(2-bromo-pyridin-4-yl)-pentanoic acid (0.12 g, 0.47 mmol) in DMF (1.5 mL), 5-bromo-pyrazin-2-ylamine (0.09 g, 0.51 mmol), 1-hydroxybenzotriazole hydrate (0.02 g, 0.17 mmol) and EDC (0.13 g, 0.70 mmol) were added. The mixture was stirred at room temperature for one hour. NaHCO₃ saturated solution was added and the mixture was extracted with DCM. The combined organic extracts were washed with saturated NaCl solution, dried over Na₂SO₄ and evaporated. The crude product was purified by silica gel chromatography (cHex/AcOEt 75/25) to give the title compound (0.02 g, 8%).

¹H NMR (400 MHz, Chloroform-d3) δ 9.59 (s, 1H), 9.29 (d, J=1.4 Hz, 1H), 8.37 (d, J=1.4 Hz, 1H), 7.57 (t, J=7.7 Hz, 1H), 7.45 (d, J=7.7 Hz, 1H), 7.27 (d, J=7.7 Hz, 1H), 3.75 (t, J=7.7 Hz, 1H), 2.21-2.16 (m, 1H), 2.08-1.92 (m, 1H), 1.45-1.21 (m, 2H), 0.95 (t, J=7.3 Hz, 3H).

C₁₅H₁₅Br₂N₃O, Calculated [414.09], found [M+H⁺] 415, RT=1.71 (method f).

Example 13: 2-(2-Bromo-pyridin-4-yl)-pentanoic Acid (5-bromo-pyridin-2-yl)-amide

(2-Bromo-pyridin-4-yl)-acetic Acid tert-butyl ester

To a solution of diisopropylamine (2.1 g, 20.93 mmol) in anhydrous THF (30 mL) under nitrogen, cooled at −78° C., a solution of n-butyllithium in hexane (2.5 M, 19.18 mmol) was added dropwise. The mixture was allowed to warm up to −30° C. and left stirring for 30 minutes. Then the reaction was cooled again to −78° C. and a solution of 2-bromo-4-methylpyridine (3.0 g, 17.44 mmol) in THF (10 mL) was added. The reaction turned to dark orange and it was stirred at −30° C. for 30 minutes. Then the reaction was cooled to −78° C. and a solution of di-tert-butyl dicarbonate (0.18 g, 19.18 mmol) in THF (10 ml) was added. Then the reaction mixture was allowed to warm up to room temperature and let stirring overnight. The mixture was quenched with H₂O and extracted with AcOEt twice. The organic layer was separated, washed with NaCl saturate solution, dried over Na₂SO₄ and concentrated under reduced pressure. The crude was purified by silica gel chromatography (cHex/AcOEt 80/20) to give the title compound (0.81 g, 13%). C₁₁H₁₄BrNO₂Mass (calculated) [272]; (found) [M+H]⁺=274.

2-(2-Bromo-pyridin-4-yl)-pentanoic Acid tert-butyl ester

The title compound was obtained following general procedure for alkylation B1 and starting from (2-bromo-pyridin-4-yl)-acetic acid tert-butyl ester (0.83 g, 3.05 mmol), (0.68 g, 71%).

C₁₄H₂0BrNO₂ Mass (calculated) [314]; (found) [M+H]⁺=316.

2-(2-Bromo-pyridin-4-yl)-pentanoic Acid

To a solution of 2-(2-Bromo-pyridin-4-yl)-pentanoic acid tert-butyl ester (0.68 g, 2.16 mmol) in DCM (20 mL), trifluoroacetic acid (2 mL) was added and the mixture was stirred at room temperature for three days. The mixture was concentrated under reduced pressure, then was diluted with DCM and extracted with NaHCO₃ saturated solution. The aqueous layer was separated, acidified to pH3 with HCl 1N and extracted with DCM. The organic phase was separated, dried over Na₂SO₄ and concentrated under reduced pressure, affording the title compound (0.44 g, 72%).

C₁₀H₁₂BrNO₂ Mass (calculated) [258]; (found) [M+H]⁺=260.

2-(2-Bromo-pyridin-4-yl)-pentanoic Acid (5-bromo-pyridin-2-yl)-amide

The title compound was obtained following the general procedure E2 for coupling with EDC and starting from 2-(2-bromo-pyridin-4-yl)-pentanoic acid and 5-bromo-pyridin-2-ylamine (0.05 g, 30%).

¹H NMR (400 MHz, Chloroform-d3) δ 8.34 (d, J=5.1 Hz, 1H), 8.31 (d, J=2.5 Hz, 1H), 8.13 (d, J=8.8 Hz, 1H), 7.89 (s, 1H), 7.81 (dd, J=8.8, 2.5 Hz, 1H), 7.52 (s, 1H), 7.28 (m, 1H), 3.42 (t, J=7.5 Hz, 1H), 2.23-2.09 (m, 1H), 1.88-1.74 (m, 1H), 1.34 (m, 2H), 0.96 (t, J=7.3 Hz, 3H).

C₁₅H₁₅Br₂N₃O, Calculated [413.11], found [M+H⁺] 414, RT=1.75 (method f).

Example 14: 2-(2-Methoxy-pyridin-4-yl)-pentanoic Acid (5-bromo-pyridin-2-yl)-amide

2-(Methoxy-pyridin-4-yl)-acetic Acid ethyl ester

The title compound was synthesized following the general procedure A1 starting from 2-methoxy-4-methyl-pyridine. (4.63 g, 73%) C₁₀H₁₃NO₃ Mass (calculated) [195]; (found) [M+H]⁺=196.

2-(2-Methoxy-pyridin-4-yl)-pentanoic Acid ethyl ester

The title compound was synthesized following the general procedure B1 starting 2-methoxy-pyridin-4-yl)-acetic acid ethyl ester. Title compound was obtained in (0.75 g, 75%).

C₁₃H₁₉NO₃ Mass (calculated) [237]; (found) [M+H]⁺=238.

2-(2-Methoxy-pyridin-4-yl)-pentanoic Acid (5-bromo-pyridin-2-yl)-amide

To 2-(2-Methoxy-pyridin-4-yl)-pentanoic acid ethyl ester (0.150 g, 0.6 mmol, 1 eq) 1,5,7-Triazabicyclo[440]dec-5-ene (0.03 g, 0.2 mmol, 0.3 eq) and 2-amino-5-bromopyridine (0.44 g, 2.5 mmol, 4 eq) were added in a vessel that was sealed with a septum and placed into the microwave cavity. Microwave irradiation (maximum emitted power 230 W) was used to increase the temperature to 130° C. The reaction mixture was then kept at this temperature for 30 min. Then the residue was diluted with DCM and washed with NaHCO₃ saturated solution. The organic phase was separated, dried over Na₂SO₄ and concentrated in vacuo. The crude was purified by silica gel chromatography (cHex/AcOEt gradient) to give the title compound (0.03 g, 15%).

¹H NMR (400 MHz, Chloroform-d3) δ 8.29-8.23 (m, 1H), 8.18-8.09 (m, 2H), 7.98 (s, 1H), 7.83-7.74 (m, 1H), 6.90-6.82 (m, 1H), 6.72 (s, 1H), 3.93 (s, 3H), 3.43 (t, J=7.5 Hz, 1H), 2.22-2.07 (m, 1H), 1.88-1.74 (m, 1H), 1.41-1.21 (m, 2H), 0.93 (t, J=7.3, 1.5 Hz, 3H).

C₁₆H₁₅BrN₃O₂, Calculated [364.24], found [M+H⁺], Br pattern 364-366, RT=1.65 (method f).

Example 15: 2-(2-Methoxy-pyridin-4-yl)-pentanoic Acid (5-chloro-thiazol-2-yl)-amide

2-(2-Methoxy-pyridin-4-yl)-pentanoic Acid

The title compound was synthesized following the general procedure D3 starting from 2-(2-methoxy-pyridin-4-yl)-pentanoic acid ethyl ester. (1.50 g, 79%).

Mass (calculated) C₁₁H₁₅NO₃ [209]; (found) [M+H⁺]=210.

Synthesis of 2-(2-Methoxy-pyridin-4-yl)-pentanoic acid (5-chloro-thiazol-2-yl)-amide

The title compound was synthesized following the general procedure E1 starting from 2-(2-methoxy-pyridin-4-yl)-pentanoic acid and 5-chloro-thiazol-2-ylamine (0.04 g, y 16%).

¹H NMR (400 MHz, Chloroform-d3) δ 10.11 (bp, 1H), 8.14 (d, J=5.4 Hz, 1H), 7.23 (s, 1H), 6.86 (dd, J=5.4, 1.4 Hz, 1H), 6.72 (d, J=1.4 Hz, 1H), 3.94 (s, 3H), 3.56 (t, J=7.5 Hz, 1H), 2.29-2.04 (m, 1H), 1.95-1.66 (m, 1H), 1.43-1.19 (m, 2H), 0.94 (t, J=7.3 Hz, 3H). C₁₄H₁₆N₃O₂SCl, Calculated [325.81], found [M+H⁺], 326, RT=2.01 (method e).

Example 16: 2-(6-Chloro-pyridin-3-yl)-pentanoic Acid (5-bromo-pyridin-2-yl)-amide

(6-Chloro-pyridin-3-yl)-acetic Acid ethyl ester

To a solution of EtOH (27 mL), concentrated H₂SO₄ (10 mL) was added dropwise and 2-chloropyridine-5-acetonitrile (2.00 g, 13.1 mmol) was added portionwise. The solution was stirred at 100° C. for three hours. The mixture was added dropwise to a solution of NaHCO₃ (30.00 g) in H₂O (100 mL) and it was extracted twice with DCM. The organic layer were collected, dried and evaporated to give the title compound (2.60 g, quant.)

C₉H₁₀ClNO₂Mass (calculated) [199]; (found) [M+H]⁺=200.

2-(6-Chloro-pyridin-3-yl)-pentanoic Acid ethyl ester

The title compound was obtained following general procedure B1 for alkylation and starting from (6-chloro-pyridin-3-yl)-acetic acid ethyl ester (0.72 g, 45%).

C₁₂H₁₆ClNO₂ Mass (calculated) [241]; (found) [M+H]⁺=242.

2-(6-Chloro-pyridin-3-yl)-pentanoic Acid

2-(6-Chloro-pyridin-3-yl)-pentanoic acid ethyl ester (0.72 g, 2.96 mmol) was dissolved in concentrated HCl (8 mL) and the solution was stirred at 100° C. for two hours. The mixture was concentrated under reduce pressure and the crude product was used in the next step without further purification (1.00 g, quant.).

C₁₀₀H₁₂ClNO₂ Mass (calculated) [213]; (found) [M+H]⁺=214.

2-(6-Chloro-pyridin-3-yl)-pentanoic Acid (5-bromo-pyridin-2-yl)-amide

The title compound was obtained following general procedure E1 for amide coupling and starting from 2-(6-Chloro-pyridin-3-yl)-pentanoic acid and 5-bromo-pyridin-2-ylamine, (0.12 g, 65%).

C₁₅H₁₅BrClN₃O Mass (calculated) [368]; (found) [M+H]⁺=370.

Example 17: 2-(5-Bromo-pyridin-3-yl)-N-(5-bromo-pyridin-2-yl)-3-(5-trifluoromethyl-furan-2-yl)-propionamide

2-(5-Bromo-pyridin-3-yl)-3-(5-trifluoromethyl-furan-2-yl)-propionic Acid ethyl ester

The title compound was obtained starting from (5-bromo-pyridin-3-yl)-acetic acid ethyl ester and 2-bromomethyl-5-methyl-furan following general procedure B1 for alkylation (0.33 g, 72%).

C₁₈H₁₃BrF₃NO₃Mass (calculated) [392]; found [M+1] 392-394 bromine pattern.

2-(5-Bromo-pyridin-3-yl)-3-(5-trifluoromethyl-furan-2-yl)-propionic Acid

The title compound was obtained using general procedure D3 for ester hydrolysis and starting from 2-(5-bromo-pyridin-3-yl)-3-(5-trifluoromethyl-furan-2-yl)-propionic acid ethyl ester (0.30 g, quant.).

C₁₃H₉BrF₃NO₃Mass (calculated) [364]; found [M+1] 364-366 bromine pattern.

2-(5-Bromo-pyridin-3-yl)-N-(5-bromo-pyridin-2-yl)-3-(5-trilfuoromethyl-furan-2-yl)-propionamide

Acid (0.06 g, 0.165 mmol, 1 eq) and 5-bromo-2-aminopyrdine (0.029 g, 0.165 mmol, 1 eq) were dissolved in AcOEt (2 mL), DIPEA (0.057 mL, 0.33 mmol, 2 eq) were added and solution cooled to 0° C. T3P 50% solution in AcOEt (0.127 mL, 0.33 mmol, 1.5 eq) was added and reaction was stirred for 12 h at room temperature. NaHCO₃ saturated solution (2 mL) was added; organic layer was separated, dried over Na₂SO₄, filtered and evaporated. The crude product was purified by silica gel chromatography (cHex/0-35% AcOEt) to give the title compound (0.08 g, 78%).

¹H NMR (400 MHz, Chloroform-d3) δ 8.63 (d, J=2.2 Hz, 1H), 8.45 (d, J=2.0 Hz, 1H), 8.30 (d, J=2.4 Hz, 1H), 8.12 (d, J=8.9 Hz, 1H), 7.97 (bs, 1H), 7.92 (t, J=2.0 Hz, 1H), 7.82 (dd, J=8.8, 2.4 Hz, 1H), 6.63 (d, J=3.3, 1H), 6.09 (d, J=3.3 Hz, 1H), 3.95 (t, J=7.7 Hz, 1H), 3.59 (m, 1H), 3.17 (m, 1H).

C₁₅H₁₂N₃O₂F₃Br₂, Calculated [519.11], found [M+H⁺], 520, RT=1.81 (method f).

Example 18: 2-(6-Methoxy-pyridin-3-yl)-pentatonic Acid (5-bromo-pyridin-2-yl)-amide

Cyano-(6-methoxy-pyridin-3-yl)-acetic Acid ethyl ester

Ethyl cyanoacetate (0.938 mL, 8.8 mmol, 1 eq) and 5-bromo-2-methoxy-pyridine (1.3 mL, 10 mmol, 1.2 eq) were added to a suspension of potassium tert-butoxide (2.4 g, 21.4 mmol, 2.5 eq) in 1,4-dioxane (25 mL) dry under N₂ atmosphere. A solution of palladium acetate (0.039 g, 0.17 mmol, 0.02 eq) and Qphos (0.198 g, 0.39 mmol, 0.04 eq) in 1,4-dioxane (10 mL) dry was added dropwise to reaction mixture. The reaction was heated at 70° C. for 2 h before cooling to room temperature; 1N acetic acid solution (15 mL) and AcOEt (20 mL) were added, organic layer was collected, dried over Na₂SO₄ and evaporated. The crude product was purified by silica gel chromatography with (cHex—10% AcOEt) to give the title compound (1.13 g, 60%).

C₁₁H₁₂N₂O₃Mass (calculated) [220]; (found) [M+H]+=221.

2-Cyano-2-(6-methoxy-pyridin-3-yl)-pentanoic Acid ethyl ester

Cyano-(6-methoxy-pyridin-3-yl)-acetic acid ethyl ester (1.13 g, 5.1 mmol, 1 eq) was dissolved in dimethylformamide (10 mL), cesium carbonate (2 g, 6.12 mmol, 1.2 eq) and 1-iodo-propane (0.55 mL, 5.6 mmol, 1.1 eq) were added and the mixture was stirred at room temperature overnight. H₂O (500 mL) was added and crude extracted three times with AcOEt (3×100 mL). Organic phases were combined, dried over Na₂SO₄ and concentrated in vacuo. The crude product was purified by silica gel chromatography (cHex-10% AcOEt) to give the title compound (1 g, 74%).

C₁₄H₁₈N₂O₃ Mass (calculated) [262]; (found) [M+H]+=263.

2-(6-Methoxy-pyridin-3-yl)-pentanoic Acid

2-cyano-2-(6-methoxy-pyridin-3-yl)-pentanoic acid ethyl ester (1 g, 3.8 mmol, 1 eq) was dissolved in methanol (7.5 mL) NaOH 2N solution (7.5 mL, 15 mmol, 4 eq) was added and mixture was stirred one hour at room temperature and at 60° C. for 2 hours. The reaction mixture was acidified to pH=5 with HCl 1 N and extracted with AcOEt (3×20 ml). Organic layers were collected, dried over Na₂SO₄ and concentrated in vacuo. The crude product was purified by silica gel chromatography (cHex-33% AcOEt) to give the title compound (0.65 g, 82%).

C₁₁H₁₅NO₃Mass (calculated) [209]; (found) [M+H]⁺=210.

2-(6-Methoxy-pyridin-3-yl)-pentatonic Acid (5-bromo-pyridin-2-yl)-amide

The amide coupling was performed using general procedure E1 to give the title compound (0.01 g, 5%).

¹H NMR (400 MHz, Chloroform-d) δ 8.42 (s, 1H), 8.26 (d, J=2.5 Hz, 1H), 8.22-8.11 (m, 2H), 7.79 (dd, J=8.9, 2.5 Hz, 1H), 7.72 (dd, J=8.7, 2.6 Hz, 1H), 6.80 (d, J=8.7 Hz, 1H), 3.97 (s, 3H), 3.58 (t, J=7.7 Hz, 1H), 2.21-2.10 (m, 1H), 1.92-1.65 (m, 1H), 1.49-1.15 (m, 2H), 0.93 (t, J=7.3 Hz, 3H).

C₁₆H₁₈N₃O₂Br, Calculated [364.24], found [M+H⁺], Br pattern 364-366, RT=1.67 (method f).

Example 19: 2-(5-Bromo-2,3-dihydro-indol-1-yl)-pentanoic Acid (5-bromo-pyridin-2-yl)-amide

2-Bromo-pentanoic Acid (5-bromo-pyridin-2-yl)-amide

2-Bromopentanoic acid (2.17 mL, 16.57 mmol, 1 eq) was dissolved in dichloroethane (15 mL) solution was reach to 0° C., Oxalyl chloride (2.90 mL, 33.15 mmol, 2 eq) was added follow by 1 drop of DMF and reaction was stirred at room temperature for 5 h. Solution was evaporated to dryness. Acyl chloride was dissolved in dichloroethane (15 mL) and slowly added to a solution of 5-bromo-2-aminopyridine (3.1 g, 18.23 mmol, 1.1 eq) and DIPEA (5.78 mL, 33.15 mmol, 2 eq) in dichloroethane over a period of 10 minutes; reaction was stirred at room temperature for 1 h. NaHCO₃ saturated solution was added, organic phase was collected, washed with a saturated solution of NaCl, dried over Na₂SO₄, filtered and evaporated. The crude product was purified by silica gel chromatography (cHex-5% AcOEt) to give the title compound (3.3 g, 65%).

C₁₀H₁₂Br₂N₂O Mass (calculated) [336]; found [M+1]=336-338 bromine pattern.

2-(5-Bromo-2,3-dihydro-indol-1-yl)-pentanoic Acid (5-bromo-pyridin-2-yl)-amide

2-Bromo-pentanoic acid (5-bromo-pyridin-2-yl)-amide (0.13 g, 0.39 mmol, 1 eq) was dissolved in CH₃CN (2 mL), DIPEA (0.081 mL, 0.46 mmol, 1.2 eq) and 5-bromoindoline (0.087 mL, 0.46 mmol, 1.2 eq) were added. Reaction was heated at 70° C. for 16 h. Acetonitrile was evaporated, the crude residue was partitioned in AcOEt (2 mL) and H₂O (2 mL), organic phase was collected, dried over Na₂SO₄, filtered and evaporated. The crude product was purified by reverse phase chromatography to give the title compound (0.021 g, 12%).

¹H NMR (400 MHz, Chloroform-d3) δ 8.97 (s, 1H), 8.29 (d, J=2.5 Hz, 1H), 8.21 (d, J=8.8 Hz, 1H), 7.81 (dd, J=8.8, 2.5 Hz, 1H), 7.22 (d, J=2.0 Hz, 1H), 7.14 (dd, J=8.4, 2.0 Hz, 1H), 6.33 (d, J=8.4 Hz, 1H), 3.95 (dd, J=7.5, 6.4 Hz, 1H), 3.62-3.43 (m, 2H), 3.16-2.97 (m, 2H), 2.23-2.00 (m, 1H), 1.79 (m, 1H), 1.55-1.29 (m, 2H), 0.96 (t, J=7.3 Hz, 3H).

C₁₅H₁₉N₃OBr₂, Calculated [453.17], found [M+H⁺], 2Br pattern 454, RT=2.12 (method f).

Example 20: 2-(5-Bromo-3-methyl-indol-1-yl)-pentanoic Acid (5-bromo-pyridin-2-yl)-amide

2-(5-Bromo-3-methyl-indol-1-yl)-pentanoic Acid ethyl ester

To a solution of 5-bromo-3-methyl-1H-indole (0.50 g, 2.38 mmol) in DMF (4 mL), NaH (60% in mineral oil, 0.11 g, 2.85 mmol) was added and the mixture was stirred at room temperature for 30 minutes. 2-Bromo-pentanoic acid ethyl ester (0.45 mL, 2.62 mmol) was added and the reaction was left stirring at room temperature overnight. Saturated NaCl solution was added and the mixture was extracted with DCM. The organic phase was collected, dried over Na₂SO₄ and concentrated under reduced pressure.

The crude product was purified by silica gel chromatography (cHex/AcOEt 80/20) to afford the title compound (0.35 g, 46%).

C₁₆H₂₀BrNO₂ Mass (calculated) [338]; (found) [M+H]⁺=340.

2-(5-Bromo-3-methyl-indol-1-yl)-pentanoic Acid (5-bromo-pyridin-2-yl)-amide

The title compound was obtained following general procedure F1 for amide coupling and starting from 2-(5-bromo-3-methyl-indol-1-yl)-pentanoic acid ethyl ester and 5-bromo-pyridin-2-ylamine (0.08 g, 34%).

¹H NMR (400 MHz, Chloroform-d) δ 8.20 (d, J=2.4 Hz, 1H), 8.15 (d, J=8.9 Hz, 1H), 7.79 (dd, J=8.9, 2.4 Hz, 1H), 7.76-7.71 (m, 2H), 7.32 (dd, J=8.7, 1.9 Hz, 1H), 7.16 (d, J=8.7 Hz, 1H), 7.01 (s, 1H), 4.90 (dd, J=10.7, 4.7 Hz, 1H), 2.48-2.36 (m, 1H), 2.34 (s, 3H), 2.23-2.07 (m, 1H), 1.33-1.12 (m, 2H), 0.93 (t, J=7.3 Hz, 3H). C₁₉H₁₉Br₂N₃O, Calculated [465.18], found [M+H⁺], 2Br pattern, 466, RT=5.38 (method c).

Example 21: 2-(3-Bromo-pyrrol-1-yl)-pentanoic Acid (5-bromo-pyridin-2-yl)-amide

2-(3-Bromo-pyrrol-1-yl)-pentanoic Acid ethyl ester

To a solution of 3-bromo-1-triisopropylsilanyl-1H-pyrrole (1.10 g, 3.64 mmol) in THF (11 mL), a solution of tetrabutyl ammonium fluoride in THF (1M, 3.82 mL, 3.82 mmol) was added and reaction was stirred for 10 minutes at room temperature. 5 mL of diethyl ether were added and the mixture was washed with 10 mL of H₂O. The organic phase was collected, dried over Na₂SO₄ and concentrated under reduced pressure, obtaining 3-bromo-1H-pyrrole that was used without further purification. To a suspension of NaH (60% in mineral oil, 0.10 g, 4.11 mL) in THF (9 mL), under nitrogen atmosphere, a solution of 3-bromo-1H-pyrrole (0.50 g, 3.46 mmol) was added and the mixture was left stirring at room temperature for 1 hour. Then the reaction was cooled down to 0° C. and a solution of 2-bromo-pentanoic acid ethyl ester (0.86 g, 4.11 mmol) in DMF (9 mL) was added. The reaction was allowed to warm up to room temperature and was left stirring for 3 hours. Then H₂O was added (10 mL) and the mixture was extracted with AcOEt (10 mL), the organic phase was collected, dried over Na₂SO₄ and concentrated under reduced pressure. The crude product was purified by silica gel chromatography (cHex/AcOEt 95/5) to afford the title compound (0.45 g, 60%).

C₁₁H₁₆BrNO₂ Mass (calculated) [274]; (found) [M+H]⁺=276.

2-(3-Bromo-pyrrol-1-yl)-pentanoic Acid (5-bromo-pyridin-2-yl)-amide

The title compound was obtained following the general procedure F1 for amide coupling and starting from 2-(3-bromo-pyrrol-1-yl)-pentanoic acid ethyl ester and 5-bromo-pyridin-2-ylamine (0.06 g, 32%).

¹H NMR (400 MHz, DMSO-d6) δ 11.00 (s, 1H), 8.45 (t, J=1.5 Hz, 1H), 8.04-7.98 (m, 2H), 6.97 (t, J=2.0 Hz, 1H), 6.86 (t, J=2.7 Hz, 1H), 6.08 (dd, J=2.7, 2.0 Hz, 1H), 4.91 (dd, J=8.9, 6.5 Hz, 1H), 2.06-1.85 (m, 2H), 1.22-1.08 (m, 2H), 0.87 (t, J=7.4 Hz, 3H).

C₁₄H₁₅Br₂N₃O, Calculated [401.10], found [M+H⁺], 2Br pattern, 402, RT=1.87 (method f).

Example 22: 2-(4-Bromo-3-trifluoromethyl-pyrazol-1-yl)-pentanoic Acid (5-bromo-pyrazin-2-yl)-amide

2-(4-Bromo-3-trifluoromethyl-pyrazol-1-yl)-pentanoic Acid ethyl ester

A suspension of 4-bromo-3-trifluoromethyl-1H-pyrazole (0.74 g, 3.44 mmol) and K₂CO₃ (0.95 g, 6.88 mmol) in acetone (16 mL) was heated at 55° C. for 10 minutes and then was allowed to cool down to room temperature. 2-Bromo-pentanoic acid ethyl ester (0.79 g, 3.78 mmol) was the added and the mixture was heated at 55° C. for 18 hours. The solvent was removed under reduced pressure and the residue was suspended in DCM and washed with H₂O. The organic phase was collected, dried over Na₂SO₄ and concentrated under reduced pressure to afford the title compound (1.38 g, quant.).

C₁₁H₁₄BrF₃N₂O₂ Mass (calculated) [343]; (found) [M+H]⁺=345.

2-(4-Bromo-3-trifluoromethyl-pyrazol-1-yl)-pentanoic Acid (5-bromo-pyrazin-2-yl)-amide

The title compound was prepared following general procedure F1 for amide coupling and starting from 2-(4-Bromo-3-trifluoromethyl-pyrazol-1-yl)-pentanoic acid ethyl ester and 5-bromo-pyrazin-2-ylamine (0.02 g, 15%).

¹H NMR (400 MHz, Chloroform-d3) δ 9.26 (s, 1H), 8.79 (s, 1H), 8.38 (s, 1H), 7.71 (s, 1H), 4.92 (t, J=7.7 Hz, 1H), 2.25 (q, J=7.7 Hz, 2H), 1.43-1.23 (m, 2H), 0.99 (t, J=7.5 Hz 3H). C₁₃H₁₂Br₂F3N₅O, Calculated [471.07], found [M+H⁺], 2Br pattern, 472. RT=1.81 (method f).

Example 23: 2-(4-Bromo-3-trifluoromethyl-pyrazol-1-yl)-pentanoic Acid (5-bromo-thiazol-2-yl)-amide

2-(4-Bromo-3-trifluoromethyl-pyrazol-1-yl)-pentanoic Acid

The title compound was prepared following general procedure D3 for ester hydrolysis and starting form 2-[4-(4-Methoxy-phenyl)-3-trifluoromethyl-pyrazol-1-yl]-pentanoic acid ethyl ester (0.50 g, quant.).

C₉H₁₀BrF₃N₂O₂ Mass (calculated) [316]; (found) [M+H]⁺=318.

2-(4-Bromo-3-trifluoromethyl-pyrazol-1-yl)-pentanoic Acid (5-bromo-thiazol-2-yl)-amide

To a solution of triphenylphosphine (0.20 g, 0.76 mmol) in DCM (2 ml) cooled at 0° C., N-bromosuccinimide (0.14 g; 0.76 mmol) was added and the mixture left at 0° C. for minutes. 2-(4-Bromo-3-trifluoromethyl-pyrazol-1-yl)-pentanoic acid (0.15 g, 0.48 mmol) was added and the reaction was allowed to warm up to room temperature and lest stirring for 45 minutes. 5-Bromo-thiazol-2-ylamine (0.31 g, 1.19 mmol) was added and the mixture was left stirring for 18 hours at room temperature. The mixture was washed with 1N HCl solution and NaHCO₃ saturated solution. The organic phase was collected and the solvent was removed under reduced pressure. The crude product was purified by silica gel chromatography (cHex/AcOEt 3/1), to afford the title compound (0.07 g, 40%).

¹H NMR (400 MHz, Chloroform-d3) δ 10.41 (s, 1H), 7.72 (s, 1H), 7.49 (s, 1H), 4.98 (dd, J=8.7, 6.8 Hz, 1H), 2.32-2.14 (m, 2H), 1.46-1.20 (m, 2H), 0.98 (t, J=7.3 Hz, 3H). C₁₂H₁₁Br₂F₃N₄OS, Calculated [476.11], found [M+H⁺], 2Br pattern, 477 RT=1.90 (method f).

Example 24: 2-(4-Bromo-3-trifluoromethyl-pyrazol-1-yl)-pentanoic Acid (3-tert-butyl-isoxazol-5-yl)-amide

The title compound was obtained following general procedure E1 for amide coupling and starting from 2-(4-Bromo-3-trifluoromethyl-pyrazol-1-yl)-pentanoic acid and 3-tert-Butyl-isoxazol-5-ylamine (0.02 g, 15%).

¹H NMR (400 MHz, Chloroform-d3) δ 9.15 (s, 1H), 7.68 (s, 1H), 6.29 (s, 1H), 4.94-4.85 (m, 1H), 2.29-2.14 (m, 2H), 1.42-1.23 (m, 11H), 0.98 (t, J=7.3 Hz, 3H).

C₁₆H₂₀N₄O₂F₃Br, Calculated [437.25], found [M+H⁺], Br pattern, 437-439, RT=1.90 (method f).

Example 25: 2-(3-Phenyl-[1,2,4]oxadiazol-5-yl)-pentanoic Acid (5-bromo-pyridin-2-yl)-amide

2-(3-Phenyl-[1,2,4]oxadiazol-5-yl)-pentanoic Acid ethyl ester

Diethyl propyl malonate (1.0 g, 4.95 mmol, 1 eq) and N-hydroxy-benzamidine (0.337 g, 2.48 mmol, 0.5 eq) were mixed in a pressure tube and heated at 140° C. for 24 h. After cooling reaction, the crude residue was dissolved in AcOEt (5 mL) and purified by silica gel chromatography (cHex-50% AcOEt) to give the title compound (0.35 g, 50%).

C₁₅H₁₈N₂O₃Mass (calculated) [274.32]; (found) [M+H]⁺=275.25.

2-(3-Phenyl-[1,2,4]oxadiazol-5-yl)-pentanoic Acid

The title compound was obtained following general procedure D3 for ester hydrolysis and starting from 2-(3-Phenyl-[1,2,4]oxadiazol-5-yl)-pentanoic acid ethyl ester, the crude product was purified by silica gel chromatography (cHex 20% AcOEt) to give the title compound (0.11 g, 30%).

C₁₃H₁₄N₂O₃Mass (calculated) [246.27]; (found) [M−H]⁻=245.3.

2-(3-Phenyl-[1,2,4]oxadiazol-5-yl)-pentanoic Acid (5-bromo-pyridin-2-yl)-amide

2-(3-Phenyl-[1,2,4]oxadiazol-5-yl)-pentanoic acid (0.11 g, 0.44 mmol, 1 eq) was dissolved in DCM (2 mL), CDI (0.798 g, 0.49 mmol, 1.1 eq) was added and reaction was stirred for 1 h at room temperature. 5-Bromo-2-aminopyridine (0.77 g, 0.44 mmol, 1 eq) was added and reaction was stirred for 16 h. NaOH 1N solution in H₂O (2 mL) was added; organic phase was collected, dried over Na₂SO₄ and concentrated under vacuo. The crude product was purified by preparative HPLC to give the title compound (0.031 g, 20%).

¹H NMR (400 MHz, Chloroform-d3) δ 9.35 (s, 1H), 8.37 (d, J=2.4 Hz, 1H), 8.20-8.06 (m, 3H), 7.82 (dd, J=8.9, 2.4 Hz, 1H), 7.60-7.48 (m, 3H), 4.13 (t, J=7.4 Hz, 1H), 2.35-2.16 (m, 2H), 1.52-1.38 (m, 2H), 0.99 (t, J=7.3 Hz, 3H).

C₁₈H₁₇N₄O₂Br, Calculated [401.26], found [M+H⁺], Br pattern 401-403, RT=1.88 (method f).

Example 26: 2-(2-Difluoromethoxy-pyridin-4-yl)-pentanoic Acid (5-bromo-pyridin-2-yl)-amide

2-(2-oxo-1,2-dihydro-pyridin-4-yl)-pentanoic Acid ethyl ester

2-(2-Methoxy-pyridin-4-yl)-pentanoic acid ethyl ester (1.0 g, 4.2 mmol, 1.0 eq.) was dissolved in acetonitrile (12 mL) at 20° C. and iodo-trimethylsilane (1.26 mL, 8.8 mmol, 2.1 eq.) was added dropwise. The mixture was heated to 80° C. for 12 h and then cooled at room temperature. The solvent was concentrated under reduced pressure and the crude product was purified by silica gel chromatography (AcOEt: cHex 1:9) to afford the title compound (0.6 g, 58%).

C₁₂H₁₇NO₃ mass (calculated) [222]; (found) [M+H]⁺=223 m/z.

2-(2-difluoromethoxy-pyridin-4-yl)-pentanoic Acid ethyl ester

2-(2-oxo-1,2-dihydro-pyridin-4-yl)-pentanoic acid ethyl ester (0.50 g, 2.2 mmol, 1.0 eq.) was dissolved in CH₃CN (10 mL) at 20° C. and sodium chloro-difluoroacetate (0.41 g, 2.7 mmol, 1.2 eq.) was added portionwise. The reaction was heated at 100° C. for 12 h and then cooled to room temperature. The solvent was distilled under reduced pressure and the crude product was purified by silica gel chromatography (AcOEt:cHex 1:9) to give the title compound (0.26 g, 42%).

C₁₃H₁₇F₂NO₃ mass (calculated) [273]; (found) [M+H]⁺=274 m/z.

2-(2-difluoromethoxy-pyridin-4-yl)-pentanoic Acid 5-(2-Br-pyridin-2-yl)-amide

2-(2-difluoromethoxy-pyridin-4-yl)-pentanoic acid ethyl ester (190 mg, 0.70 mmol, 1 eq.), 1,5,7-Triazabicyclo[4.4.0]dec-5-ene (30 mg, 0.22 mmol, 0.3 eq.) and 5-bromo-pyridin-2-yl amine (691 mg, 4 mmol, 5.7 eq.) were respectively transferred in the micro-wave tube and 2 micro-wave cycles were performed (T=130° C.; power=230 W; t=30 minutes). Then the reaction was cooled at room temperature and rinsed with dichloromethane. The organic solution was washed with sodium bicarbonate saturated solution and _(H2O). The organic layer was concentrated under reduced pressure and the crude product was purified on silica gel chromatography (AcOEt:cHex 1:5) to give the title compound (0.036 g, 13%).

¹H NMR (400 MHz, Chloroform-d3) δ 8.29 (d, J=2.5 Hz, 1H), 8.18-8.11 (m, 2H), 8.04 (s, 1H), 7.81 (dd, J=8.9, 2.5 Hz, 1H), 7.46 (t, J=73.0 Hz, 1H), 7.11 (dd, J=5.3, 1.5 Hz, 1H), 6.90 (d, J=1.5 Hz, 1H), 3.48 (t, J=7.5 Hz, 1H), 2.23-2.09 (m, 1H), 1.89-1.75 (m, 1H), 1.45-1.19 (m, 2H), 0.95 (t, J=7.3 Hz, 3H).

C₁₆H₁₆BrF₂N₃O₂, Calculated [400.2], found [M+H⁺], Br pattern, 400-402, RT=2.06 (method d).

Example 27: 2-(2-Difluoromethoxy-pyridin-4-yl)-pentanoic Acid (5-bromo-pyrazin-2-yl)-amide

2-(2-difluoromethoxy-pyridin-4-yl)-pentanoic acid ethyl ester (200 mg, 0.73 mmol, 1 eq.), 1,5,7-Triazabicyclo[4.4.0]dec-5-ene (30 mg, 0.22 mmol, 0.3 eq.) and 5-bromo-pyrazine-2-yl amine (690 mg, 4 mmol, 5.7 eq.) were respectively transferred in the micro-wave tube and 2 micro-wave cycles were performed (T=130° C.; power=230 W; t=30 minutes). Then the reaction was cooled at room temperature and rinsed with dichloromethane. The organic solution was washed with sodium bicarbonate saturated solution and H₂O. The organic layer was concentrated under reduced pressure and the crude product was purified on silica gel chromatography (AcOEt:cHex 1:5) to give the title compound (0.016 g, 7%).

¹H NMR (400 MHz, Chloroform-d3) δ 9.32 (d, J=1.4 Hz, 1H), 8.32 (d, J=1.6 Hz, 1H), 8.18 (d, J=5.3 Hz, 1H), 7.77 (s, 1H), 7.38 (t, J=72.8 Hz, 1H), 7.13 (dd, J=5.3, 1.6 Hz, 1H), 6.91 (d, J=1.4 Hz, 1H), 3.54 (t, J=7.5 Hz, 1H), 2.25-2.11 (m, 1H), 1.92-1.78 (m, 1H), 1.46-1.23 (m, 2H), 0.96 (t, J=7.3 Hz, 3H).

C₁₅H₁₅N₄O₂F₂Br, Calculated [401.21], found [M+H⁺], Br pattern, 401-403, RT=2.17 (method e).

Example 28: 22-[6-(1-Methyl-1H-pyrazol-4-yl)-pyridin-3-yl]-pentanoic Acid (5-chloro-thiazol-2-yl)-amide

2-(6-Chloro-pyridin-3-yl)-pentanenitrile

The title compound was obtained starting from 2-chloropyridine-5-acetonitrile that was treated in the same conditions of general procedure B1 for alkylation (3.80 g, 58%).

2-[6-(1-Methyl-1H-pyrazol-3-yl)-pyridin-3-yl]-pentanenitrile

The title compound was synthesized following the general procedure O for Suzuki coupling starting from 2-(6-chloro-pyridin-3-yl)-pentanenitrile and 1-methylpyrazole-4-boronic acid pinacol ester (3.80 g, 83%).

C₁₄H₁₆N₄Mass (calculated) [240]; found [M+H⁺]=241.

2-[6-(1-Methyl-1H-pyrazol-3-yl)-pyridin-3-yl]-pentanoic Acid

The starting nitrile (3.8 g, 15.6 mmol, 1 eq) was dissolved in HCl 6N aqueous solution (40 mL), solution was heated at 100° C. for 12 h. H₂O was evaporated; the solid crude product was triturated with diethyl ether, filtered off and dried to give the title compound (3.7 g, 97%).

C₁₄H₁₇N₃O₂Mass (calculated) [259]; found [M+H]⁺=260.

2-[6-(1-Methyl-1H-pyrazol-4-yl)-pyridin-3-yl]-pentanoic Acid (5-bromo-thiazol-2-yl)-amide

The title compound was obtained following procedure E1 for amide coupling with thionyl chloride after preparative HPLC purification (0.05 g, 37%).

¹H NMR (400 MHz, DMSO-d6) δ 12.65 (s, 1H), 8.44 (d, J=2.3 Hz, 1H), 8.23 (s, 1H), 7.94 (s, 1H), 7.71 (dd, J=8.3, 2.3 Hz, 1H), 7.60 (d, J=8.2 Hz, 1H), 7.50 (s, 1H), 3.90-3.81 (m, 4H), 2.11-1.97 (m, 1H), 1.80-1.66 (m, 1H), 1.28-1.13 (m, 2H), 0.87 (t, J=7.3 Hz, 3H). C₁₇H₁₈N₅OSCl, Calculated [375.88], found [M+H⁺], 376, RT=1.28 (method f).

Example 29: 2-[5-Cyano-6-(1-methyl-1H-pyrazol-4-yl)-pyridin-3-yl]-pentanoic Acid (3-tert-butyl-isoxazol-5-yl)-amide

2-(6-Chloro-5-cyano-pyridin-3-yl)-pentanoic Acid tert-butyl ester

The title compound was obtained by alkylation of (6-chloro-5-cyano-pyridin-3-yl)-acetic acid tert-butyl ester was performed following the general procedure B1 (0.60 g, 60%). C₁₅H₁₉ClN₂O₂Mass (calculated) [294]; (found) [M+H⁺]=295.

2-[5-Cyano-6-(1-methyl-1H-pyrazol-4-yl)-pyridin-3-yl]-pentanoic Acid tert-butyl ester

The title compound was synthesized following the general procedure O for Suzuki coupling, starting from 2-(6-chloro-5-cyano-pyridin-3-yl)-pentanoic acid tert-butyl ester and 1-methylpyrazole-4-boronic acid pinacol ester. The crude was purified by silica gel chromatography (cHex/AcOEt gradient) to give the title compound (0.25 g, 77%).

C₁₉H₂₄N₄O₂Mass (calculated) [340]; (found) [M+H⁺]=341.

2-[5-Cyano-6-(1-methyl-1H-pyrazol-4-yl)-pyridin-3-yl]-pentanoic Acid

The title compound was synthesized following the general procedure D2 starting from 2-[5-cyano-6-(1-methyl-1H-pyrazol-4-yl)-pyridin-3-yl]-pentanoic acid tert-butyl ester (0.20 g, quant.).

C₁₅H₁₆N₄O₂Mass (calculated)=[284]; found [M+H⁺]=285.

2-[5-Cyano-6-(1-methyl-1H-pyrazol-4-yl)-pyridin-3-yl]-pentanoic Acid (3-tert-butyl-isoxazol-5-yl)-amide

The title compound was synthesized following the general procedure E1 starting from 2-[5-cyano-6-(1-methyl-1H-pyrazol-4-yl)-pyridin-3-yl]-pentanoic acid and 3-tert-Butyl-isoxazol-5-ylamine (0.01 g, 26%).

¹H NMR (400 MHz, DMSO-d6) δ 11.86 (s, 1H), 8.72 (d, J=2.4 Hz, 1H), 8.42 (s, 1H), 8.20 (d, J=2.4 Hz, 1H), 8.11 (s, 1H), 6.23 (s, 1H), 3.92 (s, 3H), 3.89-3.78 (m, 1H), 2.11-1.97 (m, 1H), 1.83-1.69 (m, 1H), 1.28-1.14 (m, 11H), 0.87 (t, J=7.3 Hz, 3H).

C₂₂H₂₆N₆O₂, Calculated [406.48], found [M+H⁺], 407, RT=1.58 (method f).

Example 30: 2-[4-(4-Methoxy-phenyl)-3-trifluoromethyl-pyrazol-1-yl]-pentanoic Acid (5-bromo-pyrazin-2-yl)-amide

2-[4-(4-Methoxy-phenyl)-3-trifluoromethyl-pyrazol-1-yl]-pentanoic Acid ethyl ester

The title compound was prepared following general procedure O for Suzuki coupling and starting from 4-bromo-3-trifluoromethyl-1H-pyrazole and 4-methoxyphenyl boronic acid, after purification by silica gel chromatography (cHex/AcOEt 3/1) (0.09 g, 28%).

C₁₈H₂₁F₃N₂O₃ Mass (calculated) [370]; (found) [M+H]⁺=372.

2-[4-(4-Methoxy-phenyl)-3-trifluoromethyl-pyrazol-1-yl]-pentanoic Acid (5-bromo-pyrazin-2-yl)-amide

The title compound was prepared following general procedure F1 for amide coupling and starting from 2-[4-(4-methoxy-phenyl)-3-trifluoromethyl-pyrazol-1-yl]-pentanoic acid ethyl ester and 5-bromo-pyrazin-2-ylamine (0.01 g, 6%).

¹H NMR (400 MHz, Chloroform-d3) δ 9.29 (d, J=1.5 Hz, 1H), 9.21 (s, 1H), 8.38 (d, J=1.5 Hz, 1H), 7.63 (s, 1H), 7.35 (d, J=8.7 Hz, 2H), 6.95 (d, J=8.7 Hz, 2H), 4.93 (dd, J=8.6, 6.7 Hz, 1H), 3.85 (s, 3H), 2.38-2.23 (m, 2H), 1.42-1.28 (m, 2H), 1.00 (t, J=7.4 Hz, 3H). C₂₀H₁₉BrF₃N₅O₂, Calculated [498.30], found [M−H⁺], Br pattern, 496-498, RT=1.87 (method f).

Example 31: 2,2-Dicyclohexyl-N-(6-methyl-pyridin-2-yl)-acetamide

The title compound was prepared following general procedure for E1 amide coupling and starting from commercially available dicyclohexyl-acetic acid and N-(3-methyl-pyridin-2-yl)-amine (0.031 g, 8%).

1H NMR (400 MHz, cdcl3) δ 8.08 (d, J=8.3 Hz, 1H), 7.74 (s, 1H), 7.63-7.52 (m, 1H), 6.88 (d, J=7.5 Hz, 1H), 2.44 (s, 3H), 1.93-1.56 (m, 12H), 1.37-1.08 (m, 9H), 1.07-0.90 (m, 2H).

C₂₀H₃₀N₂O, Calculated [314,465], found [M+H⁺] 315, RT=5.2 (method c).

Examples 32-151 listed in table 1 below were made according to the method of column 3 and characterised by NMR (data not shown), and HPLC-MS (columns 5, 6, 7 and 8)

TABLE Found Synthesis Expected Retention MW Analytical Example Name method MW time (min) (M + 1) Purity method 32 2-(5-Bromo-pyridin-3-yl)-pentanoic acid (5- As in 368.66 1.71 370 96 f chloro-pyridin-2-yl)-amide Example 1 33 2-(5-Bromo-pyridin-3-yl)-pentanoic acid (5- As in 414.10 1.63 415 100 f bromo-pyrazin-2-yl)-amide Example 1 34 2-(5-Bromo-pyridin-3-yl)-pentanoic As in 427.13 1.87 428 90 f acid (5-bromo-6-methyl-pyridin-2-yl)-amide Example 1 35 2-(5-Bromo-pyridin-3-yl)-pentanoic acid (5- As in 374.68 1.73 376 97 f chloro-thiazol-2-yl)-amide Example 1 36 2-(5-Bromo-pyridin-3-yl)-pentanoic acid (6- As in 352.20 1.95 354 97 e fluoro-pyridin-2-yl)-amide Example 1 37 2-(5-Bromo-pyridin-3-yl)-pentanoic acid (6- As in 413.11 2.16 413 93 e bromo-pyridin-2-yl)-amide Example 1 38 2-(5-Bromo-pyridin-3-yl)-pentanoic acid (3- As in 380.28 2.19 382 100 e tert-butyl-isoxazol-5-yl)-amide Example 1 39 2-(6-Bromo-pyridin-2-yl)-pentanoic acid (5- As in 397.33 2.09 399 100 e bromo-thiazol-2-yl)-amide Example 1 40 2-(5-Bromo-pyridin-3-yl)-pentanoic acid (5- As in 358.23 1.58 359 98 f fluoro-thiazol-2-yl)-amide Example 1 41 2-(5-Bromo-pyridin-3-yl)-pentanoic acid (5- As in 475.24 2.6 476 98 e bromo-4-tert-butyl-thiazol-2-yl)-amide Example 1 42 2-(5-Bromo-pyridin-3-yl)-pentanoic acid (5- As in 433.16 1.82 434 98 f bromo-4-methyl-thiazol-2-yl)-amide Example 1 43 2-(5-Bromo-pyridin-3-yl)-pentanoic acid (5- As in 427.13 1.86 428 98 e bromo-3-methyl-pyridin-2-yl)-amide Example 1 44 2-(5-Bromo-pyridin-3-yl)-pentanoic acid (5- As in 419.14 2.15 420 95 e bromo-thiazol-2-yl)-amide Example 1 45 2-(5-Bromo-pyridin-3-yl)-pentanoic acid (5- As in 431.10 1.82 432 97 f bromo-6-fluoro-pyridin-2-yl)-amide Example 1 46 2-(5-Bromo-pyridin-3-yl)-pentanoic acid (5- As in 380.28 1.8 382 100 f tert-butyl-isoxazol-3-yl)-amide Example 1 47 2-(5-Bromo-pyridin-3-yl)-pentanoic acid (5- As in 402.21 1.77 404 97 f trifluoromethyl-pyridin-2-yl)-amide Example 1 48 2-(5-Bromo-pyridin-3-yl)-pentanoic acid (5- As in 441.16 1.94 442 100 f bromo-4,6-dimethyl-pyridin-2-yl)-amide Example 1 49 2-(5-Bromo-pyridin-3-yl)-pentanoic acid (5- As in 427.13 1.82 428 100 f bromo-4-methyl-pyridin-2-yl)-amide Example 1 50 2-(5-Bromo-pyridin-3-yl)-pentanoic acid (5- As in 369.64 1.59 371 100 f chloro-pyrazin-2-yl)-amide Example 1 51 2-(5-Bromo-pyridin-3-yl)-pentanoic acid (3- As in 392.17 1.76 394 95 f trifluoromethyl-isoxazol-5-yl)-amide Example 1 52 2-(5-Bromo-pyridin-3-yl)-2-fluoro-pentanoic As in 386.65 2.07 387 100 d acid (5-chloro-pyridin-2-yl)-amide Example 10 53 2-(5-Bromo-pyridin-3-yl)-2-fluoro-pentanoic As in 432.09 1.93 433 100 d acid (5-bromo-pyrazin-2-yl)-amide Example 10 54 2-(5-Bromo-pyridin-3-yl)-2-fluoro-pentanoic As in 445.12 2.51 446 95 e acid (5-bromo-6-methyl-pyridin-2-yl)-amide Example 10 55 2-(6-Bromo-pyridin-2-yl)-pentanoic acid (5- As in 413.11 1.81 414 95 f bromo-pyridin-2-yl)-amide Example 12 56 2-(6-Bromo-pyridin-2-yl)-pentanoic acid (5- As in 419.14 1.76 420 95 f bromo-thiazol-2-yl)-amide Example 12 57 2-(2-Bromo-pyridin-4-yl)-pentanoic acid (5- As in 414.10 1.64 415 95 f bromo-pyrazin-2-yl)-amide Example 13 58 2-(2-Bromo-pyridin-4-yl)-pentanoic acid (5- As in 368.66 1.69 370 100 f chloro-pyridin-2-yl)-amide Example 13 59 2-(5-Bromo-pyridin-3-yl)-pentanoic acid (5- As in 352.20 1.55 354 100 f fluoro-pyridin-2-yl)-amide Example 13 60 2-(2-Methoxy-pyridin-4-yl)-pentanoic acid (5- As in 370.27 1.66 372 100 f bromo-thiazol-2-yl)-amide Example 15 61 2-(2-Methoxy-pyridin-4-yl)-pentanoic acid (5- As in 365.23 1.88 365 97 e bromo-pyrazin-2-yl)-amide Example 15 62 2-(2-Methoxy-pyridin-4-yl)-pentanoic acid (3- As in 331.41 2.07 332 94 e tert-butyl-isoxazol-5-yl)-amide Example 15 63 2-(2-Methoxy-pyridin-4-yl)-pentanoic acid (5- As in 378.26 2.23 378 95 e bromo-6-methyl-pyridin-2-yl)-amide Example 15 64 2-(6-Chloro-pyridin-3-yl)-pentanoic As in 324.20 1.68 324 95 f acid (5-chloro-pyridin-2-yl)-amide Example 16 65 2-(6-Chloro-pyridin-3-yl)-pentanoic acid (5- As in 325.19 1.85 325 95 e chloro-pyrazin-2-yl)-amide Example 16 66 2-(5-Bromo-pyridin-3-yl)-N-(5-bromo- As in 520.10 1.73 521 100 f pyrazin-2-yl)-3-(5-trifluoromethyl-furan-2-yl)- Example propionamide 17 67 2-(5-Bromo-pyridin-3-yl)-N-(5-fluoro-pyridin- As in 458.20 1.67 458 100 f 2-yl)-3-(5-trifluoromethyl-furan-2-yl)- Example propionamide 17 68 2-(5-Bromo-pyridin-3-yl)-N-(5-chloro- As in 474.66 1.78 474 100 f pyridin-2-yl)-3-(5-trifluoromethyl-furan-2-yl)- Example propionamide 17 69 2-(5-Bromo-pyridin-3-yl)-N-(5-chloro- As in 475.65 1.7 475 100 f pyrazin-2-yl)-3-(5-trifluoromethyl-furan-2-yl)- Example propionamide 17 70 2-(6-Methoxy-pyridin-3-yl)-pentanoic acid (5- As in 370.27 1.67 372 100 f bromo-thiazol-2-yl)-amide Example 18 71 2-(2-Methyl-pyridin-4-yl)-pentanoic acid (5- As in 354.27 1.07 356 95 f bromo-thiazol-2-yl)-amide Example 18 72 2-(3-Trifluoromethyl-pyrazol-1-yl)-pentanoic As in 391.19 1.77 393 100 f acid (5-bromo-pyridin-2-yl)-amide Example 19 73 2-(4-Bromo-5-methyl-3-trifluoromethyl- As in 484.11 2 485 100 f pyrazol-1-yl)-pentanoic acid (5-bromo- Example pyridin-2-yl)-amide 19 74 2-(4-Bromo-3-trifluoromethyl-pyrazol-1-yl)- As in 470.08 1.91 471 95 f pentanoic acid (5-bromo-pyridin-2-yl)-amide Example 19 75 2-(4-Bromo-3-cyano-pyrazol-1-yl)-pentanoic As in 427.09 1.74 428 100 f acid (5-bromo-pyridin-2-yl)-amide Example 19 76 2-(5-Bromo-pyridin-3-yl)-hexanoic acid (5- As in 427.13 1.85 428 100 f bromo-pyridin-2-yl)-amide Example 2 77 2-(5-Bromo-pyridin-3-yl)-hexanoic acid (5- As in 382.68 1.81 384 100 f chloro-pyridin-2-yl)-amide Example 2 78 2-(5-Bromo-pyridin-3-yl)-hexanoic acid (5- As in 428.12 1.2 429 100 f bromo-pyrazin-2-yl)-amide Example 2 79 2-(5-Bromo-pyridin-3-yl)-hexanoic acid (3- As in 394.31 1.86 396 100 f tert-butyl-isoxazol-5-yl)-amide Example 2 80 2-(5-Bromo-pyridin-3-yl)-hexanoic acid (5- As in 445.12 1.92 446 100 f bromo-6-fluoro-pyridin-2-yl)-amide Example 2 81 2-(5-Bromo-pyridin-3-yl)-5-phenyl-pentanoic As in 445.74 1.81 446 98 f acid (5-chloro-pyrazin-2-yl)-amide Example 2 82 2-(5-Bromo-pyridin-3-yl)-5-phenyl-pentanoic As in 489.20 1.91 489 90 f acid (5-bromo-pyridin-2-yl)-amide Example 2 83 2-(5-Bromo-3-methyl-indol-1-yl)-pentanoic As in 420.73 5.28 421 95 c acid (5-chloro-pyridin-2-yl)-amide Example 20 84 2-(5-Bromo-3-methyl-indol-1-yl)-pentanoic As in 404.28 4.95 406 100 c acid (5-fluoro-pyridin-2-yl)-amide Example 20 85 2-(5-Bromo-indazol-1-yl)-pentanoic acid (5- As in 407.69 4.73 409 95 c chloro-pyridin-2-yl)-amide Example 20 86 2-(5-Bromo-pyrrolo[2,3-b]pyridin-1-yl)- As in 407.69 4.81 409 97 c pentanoic acid (5-chloro-pyridin-2-yl)-amide Example 20 87 2-(5-Bromo-pyrrolo[2,3-b]pyridin-1-yl)- As in 391.24 4.35 393 100 c pentanoic acid (5-fluoro-pyridin-2-yl)-amide Example 20 88 2-(3-Bromo-pyrrol-1-yl)-pentanoic acid (5- As in 356.65 1.84 358 100 f chloro-pyridin-2-yl)-amide Example 21 89 2-(3-Bromo-pyrrol-1-yl)-pentanoic acid (5- As in 402.08 1.78 403 100 f bromo-pyrazin-2-yl)-amide Example 21 90 2-(4-[4-methoxy-phenyl]-3-trifluoromethyl- As in 371.66 1.81 373 93 f pyrazol-1-yl)-pentanoic acid (5-bromo- Example pyrazin-2-yl)-amide 22 91 2-(4-Bromo-3-methyl-pyrazol-1-yl)-pentanoic As in 417.10 1.74 418 100 f acid (5-bromo-pyrazin-2-yl)-amide Example 22 92 2-(4-Bromo-imidazol-1-yl)-pentanoic acid (5- As in 402.08 1.79 402 100 f bromo-pyridin-2-yl)-amide Example 22 93 2-(4-Bromo-imidazol-1-yl)-pentanoic acid (5- As in 416.11 1.93 417 100 e bromo-6-methyl-pyridin-2-yl)-amide Example 22 94 2-[3-(4-Methoxy-phenyl)-pyrazol-1-yl]- As in 430.30 1.79 432 100 f pentanoic acid (5-bromo-pyrazin- Example 2-yl)-amide 22 95 2-(4-Bromo-3-tert-butyl-pyrazol-1-yl)- As in 459.18 2.09 460 91 f pentanoic acid (5-bromo-pyrazin-2-yl)-amide Example 22 96 2-(4-Bromo-3-cyano-pyrazol-1-yl)-pentanoic As in 428.08 1.67 427 100 f acid (5-bromo-pyrazin-2-yl)-amide Example 22 97 2-(4-Bromo-3-propyl-pyrazol-1-yl)-pentanoic As in 445.15 1.97 444 98 f acid (5-bromo-pyrazin-2-yl)-amide Example 22 98 2-(4-Bromo-3-trifluoromethyl-pyrazol-1-yl)- As in 409.18 1.79 409 100 f pentanoic acid (6-fluoro-pyridin-2-yl)-amide Example 22 99 2-(4-Chloro-3-trifluoromethyl-pyrazol-1-yl)- As in 426.62 1.8 428 92 f pentanoic acid (5-bromo-pyrazin-2-yl)-amide Example 22 100 2-(4-Chloro-3-trifluoromethyl-pyrazol-1-yl)- As in 382.17 1.78 382 97 f pentanoic acid (5-chloro-pyrazin-2-yl)-amide Example 22 101 2-(4-Chloro-3-trifluoromethyl-pyrazol-1-yl)- As in 425.63 1.88 427 98 f pentanoic acid (5-bromo-pyridin-2-yl)-amide Example 22 102 2-(4-Chloro-3-trifluoromethyl-pyrazol-1-yl)- As in 381.18 1.85 381 95 f pentanoic acid (5-chloro-pyridin-2-yl)-amide Example 22 103 2-(3-Bromo-pyrrol-1-yl)-pentanoic acid (3- As in 368.27 1.83 370 100 f tert-butyl-isoxazol-5-yl)-amide Example 23 104 2-(4-Bromo-3-trifluoromethyl-pyrazol-1-yl)- As in 426.62 1.82 428 100 f pentanoic acid (5-chloro-pyrazin-2-yl)-amide Example 23 105 2-(4-Bromo-3-trifluoromethyl-pyrazol-1-yl)- As in 409.18 1.76 411 100 f pentanoic acid (5-fluoro-pyridin-2-yl)-amide Example 23 106 1-(4-Bromo-3-trifluoromethyl-pyrazol-1-yl)- As in 468.07 1.87 469 98 f cyclobutanecarboxylic acid (5-bromo-pyridin- Example 2-yl)-amide 23 107 1-(4-Bromo-3-trifluoromethyl-pyrazol-1-yl)- As in 423.62 1.84 425 100 f cyclobutanecarboxylic acid (5-chloro-pyridin- Example 2-yl)-amide 23 108 1-(4-Bromo-3-trifluoromethyl-pyrazol-1-yl)- As in 424.60 1.75 426 100 f cyclobutanecarboxylic acid (5-chloro-pyrazin- Example 2-yl)-amide 23 109 1-(4-Chloro-3-trifluoromethyl-pyrazol-1-yl)- As in 424.60 1.77 424 95 f cyclobutanecarboxylic acid (5-bromo-pyrazin- Example 2-yl)-amide 23 110 1-(4-Chloro-3-trifluoromethyl-pyrazol-1-yl)- As in 423.62 1.86 425 98 f cyclobutanecarboxylic acid (5-bromo-pyridin- Example 2-yl)-amide 23 111 1-(4-Chloro-3-trifluoromethyl-pyrazol-1-yl)- As in 379.16 1.83 379 98 f cyclobutanecarboxylic acid (5-chloro-pyridin- Example 2-yl)-amide 23 112 1-(4-Bromo-3-cyano-pyrazol-1-yl)- As in 425.08 1.69 424 95 f cyclobutanecarboxylic acid (5-bromo-pyridin- Example 2-yl)-amide 23 113 2-(4-Bromo-3-trifluoromethyl-pyrazol-1-yl)- As in 425.63 1.86 427 100 f pentanoic acid (5-chloro-pyridin-2-yl)-amide Example 23 114 2-(4-Bromo-3-trifluoromethyl-pyrazol-1-yl)- As in 426.62 1.78 428 98 f pentanoic acid (6-chloro-pyrimidin-4-yl)- Example amide 23 115 1-(4-Bromo-3-trifluoromethyl-pyrazol-1-yl)- As in 469.05 1.78 468 95 f cyclobutanecarboxylic acid (5-bromo-pyrazin- Example 2-yl)-amide 23 116 1-(4-Bromo-3-trifluoromethyl-pyrazol-1-yl)- As in 407.16 1.73 409 95 f cyclobutanecarboxylic acid (5-fluoro-pyridin- Example 2-yl)-amide 23 117 2-[6-(1-Methyl-1H-pyrazol-4-yl)-pyridin-3- As in 428.33 1.4 430 99 f yl]-pentanoic acid (5-bromo-6-methyl-pyridin- Example 2-yl)-amide 28 118 2-[5-Cyano-6-(1-methyl-1H-pyrazol-4-yl)- As in 440.30 1.48 442 90 f pyridin-3-yl]-pentanoic acid (5-bromo- Example pyrazin-2-yl)-amide 29 119 2-[5-Fluoro-6-(1-methyl-1H-pyrazol-4-yl)- As in 388.83 1.46 389 95 f pyridin-3-yl]-pentanoic acid (5-chloro- Example pyrazin-2-yl)-amide 29 120 2-[5-Fluoro-6-(1-methyl-1H-pyrazol-4-yl)- As in 387.84 1.54 388 98 f pyridin-3-yl]-pentanoic acid (5-chloro-pyridin- Example 2-yl)-amide 29 121 2-[5-Cyano-6-(1-methyl-1H-pyrazol-4-yl)- As in 395.85 1.44 396 90 f pyridin-3-yl]-pentanoic acid (5-chloro- Example pyrazin-2-yl)-amide 29 122 2-[5-Cyano-6-(1-methyl-1H-pyrazol-4-yl)- As in 378.40 1.4 379 95 f pyridin-3-yl]-pentanoic acid (5-fluoro-pyridin- Example 2-yl)-amide 29 123 2-(5-Bromo-pyridin-3-yl)-N-(5-bromo- As in 413.11 1.9 414 100 d pyridin-2-yl)-3-methyl-butyramide Example 3 124 2-(5-Bromo-pyridin-3-yl)-N-(5-chloro- As in 368.66 1.85 369 95 d pyridin-2-yl)-3-methyl-butyramide Example 3 125 2-(5-Bromo-pyridin-3-yl)-N-(5-bromo- As in 414.10 1.77 415 100 d pyrazin-2-yl)3-methyl-butyramide Example 3 126 N-(5-Bromo-3-methyl-pyridin-2-yl)-2-(5- As in 427.13 1.83 428 96 e bromo-pyridin-3-yl)-3-methyl-butyramide Example 3 127 2-(5-Bromo-pyridin-3-yl)-N-(5-chloro-thiazol- As in 374.68 1.7 376 100 f 2-yl)-3-methyl-butyramide Example 3 128 N-(5-Bromo-3-fluoro-pyridin-2-yl)-2-(5- As in 431.10 1.5 432 97 f bromo-pyridin-3-yl)-3-methyl-butyramide Example 3 129 2-(5-Bromo-pyridin-3-yl)-N-(3-tert-butyl- As in 380.28 1.78 382 100 f isoxazol-5-yl)-3-methyl-butyramide Example 3 130 N-(5-Bromo-pyrazin-2-yl)-2,2-dicyclohexyl- As in 380.32 2.52 380-382 97 d acetamide Example 31 131 N-(5-Bromo-thiazol-2-yl)-2,2-dicyclohexyl- As in 385.36 2.6 385-387 96 d acetamide Example 31 132 2,2-Dicyclohexyl-N-(5-fluoro-pyridin-2-yl)- As in 318.43 2.42 319 100 d acetamide Example 31 133 1-(5-Bromo-pyridin-3-yl)- As in 412.08 1.5 413 93 f cyclobutanecarboxylic acid (5-bromo-pyrazin- Example 4 2-yl)-amide 134 1-(5-Bromo-pyridin-3-yl)- As in 366.64 1.57 100 366 f cyclobutanecarboxylic acid (5-chloro-pyridin- Example 4 2-yl)-amid 135 1-(5-Bromo-pyridin-3-yl)- As in 425.12 1.71 426 90 f cyclopentanecarboxylic acid (5-bromo- Example 5 pyridin-2-yl)-amide 136 1-(5-Chloro-pyridin-3-yl)- As in 367.63 1.46 367 100 f cyclobutanecarboxylic acid (5-bromo-pyrazin- Example 6 2-yl)-amide 137 1-(5-Chloro-pyridin-3-yl)- As in 366.64 1.57 364 95 f cyclobutanecarboxylic acid (5-bromo-pyridin- Example 6 2-yl)-amide 138 2-(6-Chloro-5-cyano-pyridin-3-yl)-pentanoic As in 394.65 1.63 394 100 f acid (5-bromo-pyrazin-2-yl)-amide Example 8 139 2-(6-Chloro-5-cyano-pyridin-3-yl)-pentanoic As in 393.67 1.72 393 100 f acid (5-bromo-pyridin-2-yl)-amide Example 8 140 2-(6-Chloro-5-cyano-pyridin-3-yl)-pentanoic As in 349.21 1.69 349 100 f acid (5-chloro-pyridin-2-yl)-amide Example 8 141 2-(5-Chloro-pyridin-3-yl)-pentanoic acid (5- As in 369.64 1.83 369 95 f bromo-pyrazin-2-yl)amide Example 9 142 2-(5-Chloro-pyridin-3-yl)-pentanoic acid (5- As in 325.19 1.58 323 95 f chloro-pyrazin-2-yl)-amide Example 9 143 2-(5-Chloro-pyridin-3-yl)-pentanoic acid (5- As in 368.66 1.68 368 100 f bromo-pyridin-2-yl)-amide Example 9 144 2-(5-Chloro-pyridin-3-yl)-pentanoic acid (5- As in 324.20 1.65 324 100 f chloro-pyridin-2-yl)-amide Example 9 145 2-(6-Chloro-5-methyl-pyridin-3-yl)-pentanoic As in 339.22 1.65 339 95 f acid (5-chloro-pyrazin-2-yl)-amide Example 9 146 2-(6-Chloro-5-methyl-pyridin-3-yl)-pentanoic As in 383.67 1.68 383 100 f acid (5-bromo-pyrazin-2-yl)-amide Example 9 147 2-(6-Chloro-5-methyl-pyridin-3-yl)-pentanoic As in 338.23 1.72 338 100 f acid (5-chloro-pyridin-2-yl)-amide Example 9 148 2-(6-Chloro-5-methyl-pyridin-3-yl)-pentanoic As in 382.68 1.76 383 100 f acid (5-bromo-pyridin-2-yl)-amide Example 9 149 2-(2-Chloro-pyridin-4-yl)-pentanoic acid (5- As in 369.64 1.58 371 100 f bromo-pyrazin-2-yl)-amide Example 9 150 2-(2-Chloro-pyridin-4-yl)-pentanoic acid (5- As in 324.20 1.64 324 100 f chloro-pyridin-2-yl)-amide Example 9 151 2-(2-Chloro-pyridin-4-yl)-pentanoic acid (5- As in 368.66 1.67 368 100 f bromo-pyridin-2-yl)-amide Example 9

Biological Activity

Examples 1-151 were tested in the above described cellular against CHO-S1P3 R1 cells, and show IC50 values ranging from 19 nM to 590 nM.

Example 33 was tested in the in vivo assays above described, at doses ranging 10 to 60 mg/kg showing anti-neuroinflammatory and neuroprotective activity (as et out in FIGS. 1-3) and improved cognitive functions (as set out in FIG. 4).

REFERENCES

-   Anelli, V., Bassi, R., Tettamanti, G., Viani, P. and     Riboni, L. (2005) Extracellular release of newly synthesized     sphingosine-1-phosphate by cerebellar granule cells and     astrocytes. J. Neurochem., 92, 1204-1215. -   Bajwa A, Huang L, Ye H, Dondeti K, Song S, Rosin D L, Lynch K R,     Lobo P I, Li L, Okusa M D. (2012). Dendritic cell sphingosine     1-phosphate receptor-3 regulates Th1-Th2 polarity in kidney     ischemia-reperfusion injury. J Immunol. 189(5):2584-96. -   Balthasar S, Samulin J, Ahlgren H, Bergelin N, Lundqvist M, Toescu E     C, Eggo M C, Törnquist K. (2006). Sphingosine 1-phosphate receptor     expression profile and regulation of migration in human thyroid     cancer cells. Biochem J. 398(3):547-56. -   Brinkmann V. (2009). FTY720 (fingolimod) in Multiple Sclerosis:     therapeutic effects in the immune and the central nervous system. Br     J Pharmacol. 158(5):1173-82. -   Brinkmann V. (2007). Sphingosine 1-phosphate receptors in health and     disease: mechanistic insights from gene deletion studies and reverse     pharmacology. Pharmacol Ther. 115(1):84-105. -   Bradl M, Hohlfeld R. (2003). Molecular pathogenesis of     neuroinflammation. J Neurol Neurosurg Psychiatry. 74(10):1364-70. -   Camprubí-Robles M, Mair N, Andratsch M, Benetti C, Beroukas D,     Rukwied R, Langeslag M, Proia R L, Schmelz M, Ferrer Montiel A V,     Haberberger R V, Kress M. (2013). Sphingosine-1-phosphate-induced     nociceptor excitation and ongoing pain behavior in mice and humans     is largely mediated by S1P3 receptor. J Neurosci. 33(6):2582-92. -   Casamenti F, Prosperi C, Scali C, Giovannelli L, Pepeu G. (1998).     Morphological, biochemical and behavioural changes induced by     neurotoxic and inflammatory insults to the nucleus basalis. -   Int J Dev Neurosci. 16(7-8):705-14. -   Cencetti F, Bernacchioni C, Nincheri P, Donati C, Bruni P. (2010).     Transforming growth factor-beta1 induces transdifferentiation of     myoblasts into myofibroblasts via up-regulation of sphingosine     kinase-1/S1P3 axis. Mol Biol Cell. 21(6):1111-24. -   Chen L Y, Woszczek G, Nagineni S, Logun C, Shelhamer J H. (2008).     Cytosolic phospholipase A2alpha activation induced by S1P is     mediated by the S1P3 receptor in lung epithelial cells. Am J Physiol     Lung Cell Mol Physiol. 295(2):L326-35. -   Chun J, Goetzl E J, Hla T, Igarashi Y, Lynch K R, Moolenaar W.     (2002). International Union of Pharmacology. XXXIV. Lysophospholipid     receptor nomenclature. Pharmacol Rev 54: 265-269. -   Davies L, Fassbender K, Walter S. 2013. Sphingolipids in     neuroinflammation. Handb Exp Pharmacol. (216):421-30. -   Ennaceur A, Delacour J. A new one-trial test for neurobiological     studies of memory in rats. 1: Behavioral data. (1988). Behav Brain     Res. 31(1):47-59. -   Forrest M, Sun S Y, Hajdu R, Bergstrom J, Card D, Doherty G, Hale J,     Keohane C, Meyers C, Milligan J, Mills S, Nomura N, Rosen H,     Rosenbach M, Shei G J, Singer I I, Tian M, West S, White V, Xie J,     Proia R L, Mandala S. (2004). Immune cell regulation and     cardiovascular effects of sphingosine 1-phosphate receptor agonists     in rodents are mediated via distinct receptor subtypes. J Pharmacol     Exp Ther. 309(2):758-68. -   Fischer I, Alliod C, Martinier N, Newcombe J, Brana C, Pouly S.     (2011). Sphingosine kinase 1 and sphingosine 1-phosphate receptor 3     are functionally upregulated on astrocytes under pro-inflammatory     conditions. PLoS One. 6(8):e23905. -   Foster C A, Howard L M, Schweitzer A, Persohn E, Hiestand P C,     Balatoni B, Reuschel R, Beerli C, Schwartz M, Billich A. (2007).     Brain penetration of the oral immunomodulatory drug FTY720 and its     phosphorylation in the central nervous system during experimental     autoimmune encephalomyelitis: consequences for mode of action in     multiple sclerosis. J Pharmacol Exp Ther. 323(2):469-75. -   Gude D R, Alvarez S E, Paugh S W, Mitra P, Yu J, Griffiths R,     Barbour S E, Milstien S, Spiegel S. (2008). Apoptosis induces     expression of sphingosine kinase 1 to release     sphingosine-1-phosphate as a “come-and-get-me” signal. FASEB J.     22(8):2629-38. -   Harris G L, Creason M B, Brulte G B, Herr D R. (2012). In vitro and     in vivo antagonism of a G protein-coupled receptor (S1P3) with a     novel blocking monoclonal antibody. PLoS One. 7(4):e35129. -   Ishii I, Friedman B, Ye X, Kawamura S, McGiffert C, Contos J J,     Kingsbury M A, Zhang G, Brown J H, Chun J. (2001). Selective Loss of     Sphingosine 1-Phosphate Signaling with No Obvious Phenotypic     Abnormality in Mice Lacking Its G Protein coupled Receptor,     LPB3/EDG-3. J Biol Chem. 276(36): 33697-704. -   Ishii I, Fukushima N, Ye X, Chun J. (2004). Lysophospholipid     Receptors: Signaling and Biology. Annu Rev Biochem. 73:321-54. -   Kanno T, Nishizaki T. (2011). Endogenous sphingosine 1-phosphate     regulates spontaneous glutamate release from mossy fiber terminals     via S1P(3) receptors. Life Sci. 18; 89(3-4):137-40. -   Keul P, Lucke S, von Wnuck Lipinski K, Bode C, Graler M, Heusch G,     Levkau B. (2011). Sphingosine-1-phosphate receptor 3 promotes     recruitment of monocyte/macrophages in inflammation and     atherosclerosis. Circ Res. 108(3):314-23. -   Kim E S, Kim J S, Kim S G, Hwang S, Lee C H, Moon A. (2011).     Sphingosine 1-phosphate regulates matrix metalloproteinase-9     expression and breast cell invasion through S1P3-Gαq coupling. J     Cell Sci. 1; 124(Pt 13):2220-30. -   Kono M, Mi Y, Liu Y, Sasaki T, Allende M L, Wu Y P, Yamashita T,     Proia R L. (2004). The sphingosine-1-phosphate receptors S1P1, S1P2,     and S1P3 function coordinately during embryonic angiogenesis. J Biol     Chem. 279(28):29367-73. -   Kono Y, Nishiuma T, Nishimura Y, Kotani Y, Okada T, Nakamura S,     Yokoyama M. (2007). Sphingosine kinase 1 regulates differentiation     of human and mouse lung fibroblasts mediated by TGF-beta1. Am J     Respir Cell Mol Biol. 37(4):395-404. -   Lai W Q, Melendez A J, Leung B P. (2010). Role of sphingosine kinase     and sphingosine-1-phosphate in inflammatory arthritis. World J Biol     Chem. 1(11): 321-326. -   Li C, Jiang X, Yang L, Liu X, Yue S, Li L. 2009. Involvement of     sphingosine 1-phosphate (SIP)/S1P3 signaling in cholestasis induced     liver fibrosis. Am J Pathol; 175(4): 1464-72. -   Liliom K, Guan Z, Tseng J L, Desiderio D M, Tigyi G, Watsky M A.     Growth factor-like phospholipids generated after corneal injury.     (1998). Am J Physiol. 274:C1065-C1074. -   Maceyka M, Harikumar K B, Milstien S, Spiegel S. (2012).     Sphingosine-1-phosphate signaling and its role in disease. Trends     Cell Biol. 22(1):50-60. -   Maragakis N J, Rothstein J D. (2006). Mechanisms of Disease:     astrocytes in neurodegenerative disease. Nat Clin Pract Neurol.     2(12):679-89. -   Marsolais D, Rosen H. (2009). Chemical modulators of     sphingosine-1-phosphate receptors as barrier-oriented therapeutic     molecules. Nat Rev Drug Discov. 8(4):297-307. -   Means K L, Brown J H. (2009). Cardiov Res Sphingosine-1-phosphate     receptor signalling in the heart. 82, 193-200. -   Mehta D, Konstantoulaki M, Ahmmed G U, Malik A B. (2005).     Sphingosine 1-phosphate-induced mobilization of intracellular Ca2+     mediates Rac activation and adherents junction assembly in     endothelial cells. J Biol Chem. 280:17320-17328. -   Meraz-Rios M A. Toral-Rios D, Franco-Bocanegra D, Villeda-Hernandez     J, Campos-Pena V. (2013). Inflammatory process in Alzheimer's     Disease. Front Integr Neurosci. 7(59): 1-15. -   Mettu P, Deng P, Misra U, Gawdi G, Epstein D, Rao P. (2004). Role of     lysophopholipid growth factors in the modulation of aqueous humor     outflow facility. Invest. Ophthalmol. Vis. Sci. 45:2263-2271. -   Murakami A, Takasugi H, Ohnuma S, Koide Y, Sakurai A, Takeda S,     Hasegawa T, Sasamori J, Konno T, Hayashi K, Watanabe Y, Mori K, Sato     Y, Takahashi A, Mochizuki N, Takakura N. (2010). Sphingosine     1-phosphate (SIP) regulates vascular contraction via S1P3 receptor:     investigation based on a new S1P3 receptor antagonist. Mol     Pharmacol. 77(4):704-13. -   Niessen F, Schaffner F, Furlan-Freguia C, Pawlinski R, Bhattacharjee     G, Chun J, Derian C K, Andrade-Gordon P, Rosen H, Ruf W. 2008.     Dendritic cell PAR1-S1P3 signalling couples coagulation and     inflammation. Nature. 452(7187):654-8. -   Paxinos, G., Watson, C., Pennisi, M., Topple, A., 1985. Bregma,     lambda and the interaural midpoint in stereotaxic surgery with rats     of different sex, strain and weight. J. Neurosci. Methods 13,     139-143. -   Pyne S and Pyne J N. (2000). Sphingosine 1-phosphate signalling in     mammalian cells. Biochem J. 349, 385-402. -   Rosen H, Gonzalez-Cabrera P J, Sanna M G, Brown S. (2009).     Sphingosine 1-phosphate receptor signaling. Annu Rev Biochem.     78:743-68. -   Rouach N, Pébay A, Meme W, Cordier J, Ezan P, Etienne E, Giaume C,     Tencé M. (2006). S1P inhibits gap junctions in astrocytes:     involvement of G and Rho GTPase/ROCK. Eur J Neurosci.     23(6):1453-1464. -   Scali C, Giovannini M G, Prosperi C, Bartolini L, and Pepeu G     (1997). Tacrine administration enhances extracellular acetylcholine     in vivo and restores the cognitive impairment in aged rats.     Pharmacol Res 36:463-469. -   Sanna M G, Liao J, Jo E, Alfonso C, Ahn M Y, Peterson M S, Webb B,     Lefebvre S, Chun J, Gray N, Rosen H. (2004). Sphingosine 1-Phosphate     (SIP) Receptor Subtypes S1P1 and S1P3, Respectively, Regulate     Lymphocyte Recirculation and Heart Rate. J Biol Chem.     279(14):13839-48. -   Shea B S, Tager A M. (2012). Open Rheumat J. 6(Suppl 1: M8) 123-129. -   Singleton P A, Dudek S M, Ma S F, Garcia J G. (2006).     Transactivation of sphingosine 1-phosphate receptors is essential     for vascular barrier regulation. Novel role for hyaluronan and CD44     receptor family. J Biol Chem. 281(45):34381-93. -   Spiegel S, Milstien S. (2003). Exogenous and intracellularly     generated sphingosine 1-phosphate can regulate cellular processes by     divergent pathways. Biochem Soc Trans. 31(Pt 6):1216-9. -   Stamer W D, Read A T, Sumida G M, Ethier C R. (2009).     Sphingosine-1-phosphate effects on the inner wall of Schlemm's canal     and outflow facility in perfused human eyes. Exp Eye Res.     89(6):980-8. -   Sun X, Shikata Y, Wang L, Ohmori K, Watanabe N, Wada J, Shikata K,     Birukov K G, -   Makino H, Jacobson J R, Dudek S M, Garcia J G. (2009). Enhanced     interaction between focal adhesion and adherens junction proteins:     involvement in sphingosine 1-phosphate-induced endothelial barrier     enhancement. Microvasc Res. 77:304-313. -   Takasugi N, Sasaki T, Suzuki K, Osawa S, Isshiki H, Hori Y, Shimada     N, Higo T, Yokoshima S, Fukuyama T, Lee V M, Trojanowski J Q, Tomita     T, Iwatsubo T. (2011). BACE1 activity is modulated by     cell-associated sphingosine-1-phosphate. J Neurosci. 31(18):6850-7. -   Takasugi N, Sasaki T, Ebinuma I, Osawa S, Isshiki H, Takeo K, Tomita     T, Iwatsubo T. (2013). -   FTY720/fingolimod, a sphingosine analogue, reduces amyloid-β     production in neurons. PLoS One. 8(5):e64050. -   Takuwa N, Ohkura S, Takashima S, Ohtani K, Okamoto Y, Tanaka T,     Hirano K, Usui S, Wang F, Du W, Yoshioka K, Banno Y, Sasaki M, Ichi     I, Okamura M, Sugimoto N, Mizugishi K, Nakanuma Y, Ishii I, Takamura     M, Kaneko S, Kojo S, Satouchi K, Mitumori K, Chun J, Takuwa Y.     (2010). S1P3-mediated cardiac fibrosis in sphingosine kinase 1     transgenic mice involves reactive oxygen species. Cardiovasc Res.     85(3):484-93. -   Taniguchi M, Kitatani K, Kondo T, Hashimoto-Nishimura M, Asano S,     Hayashi A, Mitsutake S, Igarashi Y, Umehara H, Takeya H, Kigawa J,     Okazaki T (2012). Regulation of autophagy and its associated cell     death by “sphingolipid rheostat”: reciprocal role of ceramide and     sphingosine 1-phosphate in the mammalian target of rapamycin     pathway. J Biol Chem. 287(47):39898-910. -   Trifilieff A, Fozard J R. (2012). Sphingosine-1-phosphate-induced     airway hyper-reactivity in rodents is mediated by the     sphingosine-1-phosphate type 3 receptor. J Pharmacol Exp Ther.     342(2):399-406. -   Uhlig S, Yang Y. (2013). Sphingolipids in Disease. Handbook of     Experimental Pharmacology, Sphingolipids in Acute Lung Injury. Vol.     216, pp 227-246. -   Welch S P, Sim-Selley L J, Selley D E. (2012).     Sphingosine-1-phosphate receptors as emerging targets for treatment     of pain. Biochem Pharmacol. 84(12):1551-62. -   Wu Y P, Mizugishi K, Bektas M, Sandhoff R, Proia R L. (2008).     Sphingosine kinase 1/S1P receptor signaling axis controls glial     proliferation in mice with Sandhoff disease. Hum Mol Genet.     17(15):2257-64. -   Yamashita H, Kitayama J, Shida D, Yamaguchi H, Mori K, Osada M, Aoki     S, Yatomi Y, Takuwa Y, Nagawa H. (2006). Sphingosine 1-phosphate     receptor expression profile in human gastric cancer cells:     differential regulation on the migration and proliferation. J Surg     Res. 130(1):80-7. -   Yin Z, Fan L, Wei L, Gao H, Zhang R, Tao L, Cao F, Wang H. (2012).     FTY720 protects cardiac microvessels of diabetes: a critical role of     S1P1/3 in diabetic heart disease. PLoS One. 7(8):e42900. -   Young N and Van Brocklyn J R. (2007). Roles of     Sphingosine-1-Phosphate (S1P) Receptors in Malignant behavior of     Glioma Cells. Differential Effects of S1P2 on Cell Migration and     Invasiveness. Exp Cell Res. 1; 313(8): 1615-1627. -   Zhang Y H, Fehrenbacher J C, Vasko M R, Nicoll G D. (2006).     Sphingosine-1-Phosphate Via Activation of a G-Protein-Coupled     Receptor(s) Enhances the Excitability of Rat Sensory Neurons. J     Neurophysiol. 96: 1042-1052. -   Brinkmann V. (2007). Sphingosine 1-phosphate receptors in health and     disease: mechanistic insights from gene deletion studies and reverse     pharmacology. Pharmacol Ther. 115(1):84-105. -   Spiegel S, Milstien S. (2003). Exogenous and intracellularly     generated sphingosine 1-phosphate can regulate cellular processes by     divergent pathways. Biochem Soc Trans. 31(Pt 6):1216-9. -   Forrest M, Sun S Y, Hajdu R, Bergstrom J, Card D, Doherty G, Hale J,     Keohane C, Meyers C, Milligan J, Mills S, Nomura N, Rosen H,     Rosenbach M, Shei G J, Singer I I, Tian M, West S, White V, Xie J,     Proia R L, Mandala S. (2004). Immune cell regulation and     cardiovascular effects of sphingosine 1-phosphate receptor agonists     in rodents are mediated via distinct receptor subtypes. J Pharmacol     Exp Ther. 309(2):758-68. -   Liliom K, Guan Z, Tseng J L, Desiderio D M, Tigyi G, Watsky M A.     Growth factor-like phospholipids generated after corneal injury.     (1998). Am J Physiol. 274:C1065-C1074. -   Mettu P, Deng P, Misra U, Gawdi G, Epstein D, Rao P. (2004). Role of     lysophopholipid growth factors in the modulation of aqueous humor     outflow facility. Invest. Ophthalmol. Vis. Sci. 45:2263-2271. -   Rouach N, Pébay A, Meme W, Cordier J, Ezan P, Etienne E, Giaume C,     Tencé M. (2006). S1P inhibits gap junctions in astrocytes:     involvement of G and Rho GTPase/ROCK. Eur J Neurosci.     23(6):1453-1464. -   Casamenti F, Prosperi C, Scali C, Giovannelli L, Pepeu G. (1998).     Morphological, biochemical and behavioural changes induced by     neurotoxic and inflammatory insults to the nucleus basalis. -   Int J Dev Neurosci. 16(7-8):705-14. 

1. A compound of formula (A),

wherein ●—R₁ is

X₁, X₆, X₇, X₉ and X₁₀ are halogen, C₁-C₄ linear branched or cyclic alkyl optionally substituted with one or more fluorine atoms; X₂, X₃, X₄, X₅ and X₈, are hydrogen, halogen, C₁-C₄ linear branched or cyclic alkyl optionally substituted with one or more fluorine atoms; with the proviso that at least one of X₂, X₃, X₄, and X₅ is not hydrogen R₂ is a C₃-C₆ linear branched or cyclic alkyl optionally substituted with phenyl, with one or more fluorine atoms or with trifluoromethyl-furanyl; R₂′ is hydrogen, F, C₁-C₃ linear or branched alkyl optionally substituted with one or more fluorine atoms; or R₂ and R₂′ together with the carbon atom they are attached to form a C₃-C₆ cycloalkyl ring; ●—R₃ is

Y₁ is halogen; Y₁′ is C₁-C₃ linear branched or cyclic alkyl optionally substituted with one or more fluorine atoms; Y₂ is cyano or methoxyphenyl, C₁-C₃ linear branched or cyclic alkyl optionally substituted with one or more fluorine atoms; Y₃ is hydrogen, halogen or methoxyphenyl; Y₄ is hydrogen, halogen, N-methylpyrazolyl or a C₁-C₃ linear branched or cyclic alkoxy optionally substituted with one or more fluorine atoms, Y₅ is hydrogen halogen, cyano, or a C₁-C₃ linear branched or cyclic alkyl optionally substituted with one or more fluorine atoms; with the proviso that at least one of Y₄ and Y₅ is not hydrogen; Y₆ is halogen, C₁-C₃ linear branched or cyclic alkyl optionally substituted with one or more fluorine atoms, or a C₁-C₃ linear branched or cyclic alkoxy optionally substituted with one or more fluorine atoms; enantiomers, enantiomerically enriched mixtures, and pharmaceutically acceptable salts thereof.
 2. The compound of claim 1 wherein X₁ is halogen; X₂ is hydrogen, halogen or methyl; X₃ is hydrogen, halogen or trifluoromethyl; X₄ is hydrogen or methyl; X₅ is hydrogen or halogen; X₆ is halogen; with the proviso that at least one of X₂, X₃, X₄, and X₅ is not hydrogen; X₇ is t-butyl or trifluoromethyl, preferably t-butyl; X₈ is hydrogen, methyl or t-butyl; X₉ is halogen; X₁₀ is t-butyl; R₂ is n-propyl, 3-phenyl-n-propyl, i-propyl, n-butyl, cyclohexyl, (5-trifluoromethyl-furan-2yl)-methyl; R₂′ is hydrogen, F, methyl; or R₂ and R₂′ together with the carbon atom they are attached to form a cyclobutyl or cyclopentyl group; Y₁ is halogen; Y₁′ is methyl; Y₂ is methyl, n-propyl, cyano, trifluoromethyl or 4-methoxyphenyl; Y₃ is hydrogen, halogen, or 4-methoxyphenyl; Y₄ is hydrogen, halogen, methoxy or 1-methyl-pyrazol-4-yl; Y₅ is hydrogen, halogen, cyano or methyl; with the proviso that at least one of Y₄ and Y₅ is not hydrogen; Y₆ halogen, methoxy or difluoromethoxy.
 3. The compound of claim 1 or 2, wherein ●-R₁ is selected from

and wherein R₂, R₂′, R₃, X₁, X₂, X₃, X₄, X₅, X₈, X₉, Y₁, Y₁′, Y₂, Y₃, Y₄, Y₅ and Y₆ are as defined in claim 1 or
 2. 4. The compound of claim 3, which is selected from the group consisting of 1-(5-Chloro-pyridin-3-yl)-cyclobutanecarboxylic acid (5-chloro-pyridin-2-yl)-amide; 2-(6-Chloro-5-cyano-pyridin-3-yl)-pentanoic acid (5-chloro-pyrazin-2-yl)-amide; 2-(6-Chloro-5-fluoro-pyridin-3-yl)-pentanoic acid (5-bromo-pyrazin-2-yl)-amide; 2-(6-Bromo-pyridin-2-yl)-pentanoic acid (5-bromo-pyrazin-2-yl)-amide; 2-(2-Bromo-pyridin-4-yl)-pentanoic acid (5-bromo-pyridin-2-yl)-amide; 2-(2-Methoxy-pyridin-4-yl)-pentanoic acid (5-bromo-pyridin-2-yl)-amide; 2-(6-Methoxy-pyridin-3-yl)-pentanoic acid (5-bromo-pyridin-2-yl)-amide; 2-(4-Bromo-3-trifluoromethyl-pyrazol-1-yl)-pentanoic acid (5-bromo-pyrazin-2-yl)-amide; 2-(2-Difluoromethoxy-pyridin-4-yl)-pentanoic acid (5-bromo-pyridin-2-yl)-amide; 2-[6-(1-Methyl-1H-pyrazol-4-yl)-pyridin-3-yl]-pentanoic acid (5-chloro-thiazol-2-yl)-amide; 2-(5-Bromo-pyridin-3-yl)-pentanoic acid (5-bromo-pyrazin-2-yl)-amide; 2-(5-Bromo-pyridin-3-yl)-pentanoic acid (5-bromo-3-methyl-pyridin-2-yl)-amide; 2-(5-Bromo-pyridin-3-yl)-pentanoic acid (5-bromo-6-fluoro-pyridin-2-yl)-amide; 2-(5-Bromo-pyridin-3-yl)-pentanoic acid (5-chloro-pyrazin-2-yl)-amide; 2-(5-Bromo-pyridin-3-yl)-2-fluoro-pentanoic acid (5-chloro-pyridin-2-yl)-amide; 2-(2-Bromo-pyridin-4-yl)-pentanoic acid (5-bromo-pyrazin-2-yl)-amide; 2-(2-Bromo-pyridin-4-yl)-pentanoic acid (5-chloro-pyridin-2-yl)-amide; 2-(5-Bromo-pyridin-3-yl)-pentanoic acid (5-fluoro-pyridin-2-yl)-amide; 2-(2-Methoxy-pyridin-4-yl)-pentanoic acid (5-bromo-thiazol-2-yl)-amide; 2-(2-Methoxy-pyridin-4-yl)-pentanoic acid (5-bromo-pyrazin-2-yl)-amide; 2-(4-Bromo-3-trifluoromethyl-pyrazol-1-yl)-pentanoic acid (5-bromo-pyridin-2-yl)-amide; 2-(4-Bromo-3-cyano-pyrazol-1-yl)-pentanoic acid (5-bromo-pyridin-2-yl)-amide; 2-(5-Bromo-pyridin-3-yl)-hexanoic acid (5-bromo-pyrazin-2-yl)-amide; 2-(4-[4-methoxy-phenyl]-3-trifluoromethyl-pyrazol-1-yl)-pentanoic acid (5-bromo-pyrazin-2-yl)-amide; 2-(4-Bromo-3-methyl-pyrazol-1-yl)-pentanoic acid (5-bromo-pyrazin-2-yl)-amide; 2-(4-Bromo-imidazol-1-yl)-pentanoic acid (5-bromo-pyridin-2-yl)-amide; 2-[3-(4-Methoxy-phenyl)-pyrazol-1-yl]-pentanoic acid (5-bromo-pyrazin-2-yl)-amide; 2-(4-Bromo-3-cyano-pyrazol-1-yl)-pentanoic acid (5-bromo-pyrazin-2-yl)-amide; 2-[5-Fluoro-6-(1-methyl-1H-pyrazol-4-yl)-pyridin-3-yl]-pentanoic acid (5-chloro-pyrazin-2-yl)-amide; 2-[5-Cyano-6-(1-methyl-1H-pyrazol-4-yl)-pyridin-3-yl]-pentanoic acid (5-chloro-pyrazin-2-yl)-amide; 2-(5-Bromo-pyridin-3-yl)-N-(5-bromo-pyrazin-2-yl)3-methyl-butyramide; N-(5-Bromo-3-fluoro-pyridin-2-yl)-2-(5-bromo-pyridin-3-yl)-3-methyl-butyramide; N-(5-Bromo-pyrazin-2-yl)-2,2-dicyclohexyl-acetamide; 1-(5-Bromo-pyridin-3-yl)-cyclobutanecarboxylic acid (5-bromo-pyrazin-2-yl)-amide; 1-(5-Chloro-pyridin-3-yl)-cyclobutanecarboxylic acid (5-bromo-pyrazin-2-yl)-amide; 2-(6-Chloro-5-cyano-pyridin-3-yl)-pentanoic acid (5-bromo-pyrazin-2-yl)-amide; 2-(5-Chloro-pyridin-3-yl)-pentanoic acid (5-bromo-pyrazin-2-yl)amide; 2-(5-Chloro-pyridin-3-yl)-pentanoic acid (5-chloro-pyrazin-2-yl)-amide; 2-(6-Chloro-5-methyl-pyridin-3-yl)-pentanoic acid (5-chloro-pyrazin-2-yl)-amide; and 2-(2-Chloro-pyridin-4-yl)-pentanoic acid (5-bromo-pyrazin-2-yl)-amide.
 5. The compound of claim 1 or 2 wherein ●-R₁ is

and wherein R₂, R₂′, R₃, X₇, Y₁, Y₁′, Y₂, Y₃, Y₄, Y₅ and Y₆ are as defined in claim 1 or
 2. 6. The compound of claim 5, which is selected from the group consisting of 2-(5-Bromo-pyridin-3-yl)-pentanoic acid (3-tert-butyl-isoxazol-5-yl)-amide and 2-(5-Bromo-pyridin-3-yl)-hexanoic acid (3-tert-butyl-isoxazol-5-yl)-amide.
 7. The compound of claim 1 or 2, wherein ●-R₃ is selected from

and wherein R₁, R₂, R₂′, X₁, X₂, X₃, X₄, X₅, X₆, X₇, X₈, X₉, X₁₀, Y₂, Y₃, Y₄, Y₅ and Y₆ are as defined in claim 1 or
 2. 8. The compound of claim 3 or 5, wherein ●-R₃ is selected from

and wherein R₁, R₂, R₂′, X₁, X₂, X₃, X₄, X₅, X₆, X₇, X₈, X₉, X₁₀, Y₂, Y₃, Y₄, Y₅ and Y₆ are as defined in claim 3 or
 5. 9. A pharmaceutical composition containing a compound according to claims 1-8.
 10. The compound of claims 1-8 for use as medicaments.
 11. The compound of claim 10 for use in the treatment of arthritis, fibrosis, inflammatory syndromes, atherosclerosis, vascular diseases, asthma, bradycardia, acute lung injury, lung inflammation, cancer, ocular hypertension, glaucoma, neuroinflammatory diseases, neurodegenerative diseases, Sandhoff s disease, kidney ischemia-reperfusion injury, pain, diabetic heart disease.
 12. The compound of claim 10 for use in the treatment of Alzheimer's disease, Parkinson's disease, Amyotrophic lateral sclerosis, Huntington's disease and Multiple Sclerosis.
 13. The compounds of claim 10 for use in the treatment of Alzheimer's disease. 